Comparison of different methods of sputum aspiration in children with bronchopneumonia / 中国实用护理杂志

Objective:To compare the effect of different methods of sputum aspiration in children with bronchopneumonia and explore the appropriate sputum aspiration methods for children with bronchopneumonia.Methods:This study was a randomized controlled trial. The convenience sampling method was used to select 165 children with bronchopneumonia treated in the Third Affiliated Hospital of Zhengzhou University from October 8, 2020 to September 30, 2021 as the research objects. They were divided into group A, group B and group C by random number table method, with 55 cases in each group. Group A used sputum aspirating tube to aspirate sputum through mouth and nose, group B used olive head to aspirate sputum through nose, and group C used both sputum aspirating tube to aspirate sputum through mouth and olive head through nose. The sputum aspirating effect, respiratory mucosal injury rate and satisfaction degree of children′s parents were compared among the three groups.Results:Finally, there were 53 cases in group A, 54 cases in group B and 53 cases in group C. The total effective rate of sputum aspiration was 96.21%(584/607) in group C, which was higher than that in group A 94.32% (581/616) and in group B 74.12% (464/626), the difference was statistically significant ( χ2=194.62, P<0.01), pairwise comparison among the three groups showed statistical difference ( χ2=95.95, 11.32, 132.16, all P<0.017). The respiratory mucosal injury rate was 1.12% (7/626) in group B, which was lower than that in group C 3.13% (19/607) and in group A 10.39% (64/616), the difference was statistically significant ( χ2=63.53, P<0.01), pairwise comparison among the three groups showed statistical difference ( χ2=49.52, 25.47, 6.04, all P<0.017). The satisfaction degree of children′s parents in group C was 98.11% (52/53), which was higher than that in group B 90.74% (49/54) and in group A 81.13% (43/53), the difference was statistically significant ( χ2=8.54, P<0.05). There was no statistically significant difference in the satisfaction degree of children′s parents between group A and group B, and between group B and group C( P>0.017). There was statistically significant difference in the satisfaction degree of children′s parents between group A and group C ( χ2=8.22, P<0.017). Conclusions:The combination of oral suction tube and nasal suction with olive head can improve the sputum aspirating effect of children with bronchopneumonia and the satisfaction degree of their parents, with a low rate of respiratory mucosal injury and high safety, which is worthy of clinical reference.

Effect of Maxing Shigantang and Supplemented Guominjian Decoction on Symptoms and Inflammatory Cytokine Levels in Induced Sputum of Children with Cough Variant Asthma / 中国实验方剂学杂志

ObjectiveTo investigate effect of Maxing Shigantang and supplemented Guominjian decoction on symptoms and levels of inflammatory cytokines in induced sputum of children with cough variant asthma （CVA）. MethodA total of 118 CVA children who were treated in our hospital from January 2020 to January 2021 were enrolled and randomized into the control group and the observation group with the random number table method. Control group received routine western medicine and the observation group was treated by routine western medicine， Maxing Shigantang， and supplemented Guominjian decoction. In the one-month follow-up， the scores of cough and accompanying symptoms， levels of inflammatory cytokines ［interleukin-10 （IL-10）， interleukin-5 （IL-5）， tumor necrosis factor-α （TNF-α）， neutrophil， eosinophil］ in induced sputum， pulmonary function parameters ［forced vital capacity （FVC）， forced expiratory volume in one second （FEV1）， FEV1/FVC］， and treatment outcomes were compared between the two groups. Moreover， the frequency of acute asthma attacks during the three-month follow-up was also compared. ResultNo cases dropped out from this study. After treatment， the scores of cough and accompanying symptoms were decreased in both groups （P<0.05） and were lower in observation group than in control group （P<0.05）. After treatment， FVC， FEV1， and FEV1/FVC were raised in both groups and were higher in observation group than in control group （P<0.05）. The increase in the level of IL-10 along with the decrease in levels of IL-5， TNF-α， neutrophil， and eosinophil in induced sputum was found in both groups after treatment （P<0.05）， and observation group had higher level of IL-10 and lower levels of IL-5， TNF-α， neutrophil， and eosinophil than the control group （P<0.05）. The effective rate was 86.44% （51/59） in observation group， which was higher than the 69.49% （41/59） in control group （χ2=4.933， P<0.05）. No serious adverse reaction occurred in either group. The frequency of acute asthma attacks during the three-month follow-up was （1.09±0.18） in observation group， which was lower than the （2.83±049） in the control group （P<0.05）. ConclusionRoutine western medicine combined with Maxing Shigantang and supplemented Guominjian decoction can effectively and safely alleviate the airway inflammatory responses， control the clinical symptoms， improve pulmonary function， and reduce the frequency of acute recurrence in the treatment of CVA children.

Development and validity of an educational folder for pulmonary tuberculosis sputum collection / Desarrollo y validación de una carpeta educativa para la recolección de esputo de tuberculosis pulmonar / Desenvolvimento e validação de um folder educativo para coleta de escarro da tuberculose pulmonar

ABSTRACT Objective: to develop and validate an educational folder for pulmonary tuberculosis sputum collection. Method: a methodological study, developed in four phases: project design, bibliographic survey, development, and content and appearance validity. For validity, the Content Validity Index greater than or equal to 0.80, the Kappa Coefficient and the Suitability Assessment of Materials were adopted. Results: an educational folder was developed containing the recommended steps for quality sputum collection. It was validated by 19 expert judges in the first validity cycle, obtaining an overall Content Validity Index of 0.90, perfect agreement among judges, with a total Kappa of 0.83 and superior suitability. Conclusion: the material is valid, as it contains suitable, simple and attractive language, illustrations and layout, indicating perfect agreement and reliability. Thus, it has the potential to assist in carrying out the recommended steps for correct sputum collection and enable the correct diagnosis.

RESUMEN Objetivo: desarrollar y validar una carpeta educativa para la recolección de esputo de tuberculosis pulmonar. Método: estudio metodológico, desarrollado en cuatro fases: diseño del proyecto, levantamiento bibliográfico, elaboración y validación de contenido y apariencia. Para la validación se adoptó el Índice de Validez de Contenido mayor o igual a 0,80, el Coeficiente Kappa y el Suitability Assessment of Materials. Resultados: se elaboró una carpeta educativa con los pasos recomendados para la recolección de esputo de calidad. Fue validado por 19 jueces expertos en el primer ciclo de validación, obteniendo un Índice de Validez de Contenido global de 0,90, perfecta concordancia entre los jueces, con un Kappa total de 0,83 y adecuación superior. Conclusión: el material es válido por contener lenguaje, ilustraciones y maquetación adecuados, sencillos y atractivos, indicando perfecta concordancia y fiabilidad. Por lo tanto, tiene el potencial de ayudar a llevar a cabo los pasos recomendados para la recolección correcta de esputo y permitir el diagnóstico correcto.

RESUMO Objetivo: desenvolver e validar um folder educativo para coleta de escarro da tuberculose pulmonar. Método: estudo metodológico, desenvolvido em quatro fases: elaboração do projeto, levantamento bibliográfico, desenvolvimento e validação de conteúdo e aparência. Para validação, adotaram-se o Índice de Validade de Conteúdo maior ou igual a 0,80, o Coeficiente Kappa e o Suitability Assessment of Materials. Resultados: desenvolveu-se um folder educativo contendo os passos recomendados para coleta de escarro de qualidade. Esse foi validado, por 19 juízes especialistas, no primeiro ciclo de validação, ao obter Índice de Validade de Conteúdo global de 0,90, concordância perfeita entre os juízes, com Kappa total de 0,83 e adequação superior. Conclusão: o material é válido, pois contém linguagem, ilustrações e layout adequados, simples e atraentes, indicando concordância e confiabilidade perfeita. Assim, apresenta potencialidade para auxiliar na realização dos passos recomendados para correta coleta de escarro e possibilitar o diagnóstico correto.

Delayed SARS-COV-2 clearance in infected persons in Ghana

Objective: This study aimed to determine the duration of SARS-CoV-2 clearance in persons in Ghana. The research question was whether the duration of virus clearance in Ghana matched the 14 days recommended by the World Health Organization (WHO); this had direct implications for transmission, which was key in managing the COVID-19 pandemic. Design: This was a retrospective analytical study. Setting: All facilities that submitted clinical specimens to Noguchi Memorial Institute for Medical Research (NMIMR) for SARS-CoV-2 diagnosis between March to June 2020 were included in the study. Interventions: Samples from 480 persons who tested positive for SARS-CoV-2 by RT-PCR from March to June 2020 at NMIMR and submitted at least two follow-up samples were retrospectively analysed. Individuals with two consecutive negative RT-PCR retesting results were considered to have cleared SARS-CoV-2. Results: The median time from the initial positive test to virus clearance was 20 days (IQR: 5-56 days). This was six days longer than the WHO-recommended 14 days, after which infected persons could be de-isolated. Sputum and nasopharyngeal swabs proved more sensitive for detecting viral RNA as the infection progressed. At a significance level of 0.05, age and sex did not seem to influence the time to SARS-CoV-2 clearance. Conclusions: The median time to SARS-CoV-2 clearance in this study was 20 days, suggesting that SARS-CoV-2 infected persons in Ghana take longer to clear the virus. This finding calls for further investigations into whether patients who remain PCR positive continue to be infectious and inform isolation practices in Ghana.

Differential expression of sigma factor of Mycobacterium tuberculosis under in vivo infection and in vitro culture conditions / 公共卫生与预防医学

Objective To investigate the gene expression of sigma factors in vivo, and to explore the sigma factors that may be closely related to the virulence of pathogenic Mycobacterium tuberculosis Methods Tuberculosis (TB) patients diagnosed in the outpatient department of Tianjin Tuberculosis Control Center from January to December 2018 were selected, and 20 sputum-positive specimens were randomly selected from TB patients confirmed with Xpert-positive for the present study. Two immediate sputum specimens were collected from each case of pulmonary tuberculosis before treatment, one for RNA extraction and one for in vitro culture. In vitro cultured strains in the logarithmic phase of growth were harvested for RNA extraction. The specific primers for 13 sigma factors were designed. The differential expression of the 13 sigma factors between sputum isolates and in vitro cultured strains was analyzed by fluorescence quantitative PCR. Taking ribosomal 16s as the reference gene, the transcription level of sigma factors was analyzed by 2ΔCt. Using the stably expressed sigA as the control reference, the expression differences of other sigma factors were analyzed by one-way ANOVA. Results Within 0 days, stress-associated sigma factors have a different expression profile in clinical isolate strains vs H37Rv or in vitro. All the sigma factors induced up regulation in sputum ,while no difference transcription between clinical isolate strains vs H37Rv（P＞0.05）. When compared to in vitro culture ,only sigM transcript highest in sputum(P＜0.05). Conclusion SigM plays an important role in the initial stages of bacterial infection, but its exact role is unclear.We assumed it could have a role in the interplay between the host immune defenses and the bacterial escape mechanisms.

Comparison between the Mycob.T scanner system and manual microscopy in detecting acid-fast bacilli in sputum specimens / 上海预防医学

ObjectiveTo compare the efficacy between the Mycob.T scanner system and manual microscopy for detecting acid-fast bacilli in sputum specimens. MethodsBetween January and November 2020， a total of 1 519 sputum samples from suspected primary tuberculosis patients from 5 designated tuberculosis hospitals in Shanghai were examined by Smear and BACTEC MGIT 960 liquid culture （liquid culture） methods. Each specimen was subiected to 2 direct smear slides. One slide was stained by Z-N method and examined with manual microscopic method. Another slide was stained and scanned by the Mycob.T system. The efficacy of manual microscopy and the Mycob.T scanner system for detecting acid-fast bacilli in sputum specimens was compared based on the result of liquid culture. Results of the repetitive scanning by the Mycob.T scanner system and the recheck of the manual microscopy were analyzed. ResultsThe average positive rate by the Mycob.T scanner system was 14.4% （219/1 519） while the average positive rate by manual microscopy was 16.3% （248/1 519）. No significant difference was observed （χ2=2.13， P=0.145）. Based on liquid culture confirmation results， the sensitivity of manual microscopy （60.36%） was higher than that of the Mycob.T scanner system （52.94%）， and the difference is statistically significant （χ2=4.38， P=0.036）. Both methods had high specificity （98.94%）. The concordance of the Mycob.T scanner system and manual microscopy was 95.46%， with the kappa value of 0.826. The results of repeatability test of the Mycob.T scanner system and the recheck results of the manual microscopy showed that the coincidence rate of scanning by the Mycob.T scanner system was 99.5% （436/438）， and the recheck coincidence rate by the manual microscopy was 98.6% （432/438）. ConclusionThe Mycob.T scanner system have high specificity for detecting acid-fast bacilli in sputum samples and good consistency with the results of manual microscopy. Compared with manual microscopic examination， the Mycob.T scanner system can greatly alleviate the work intensity.

Design and application of an extension tube for ventilator pipeline / 中华危重病急救医学

At present, the extension tube of the ventilator, a long corrugated tube used in clinical practice, needs to be disconnected from the endotracheal tube when performing open sputum suction. Under positive pressure, the condensed water containing pathogenic microorganisms in the ventilator will be splashed. Some will form aerosol suspended in the air, pollute the air, increase the risk of cross-infection and operator pollution, while the ventilator will emit a corresponding alarm sound, resulting in noise pollution. Based on the above factors, a ventilator pipeline extension tube was designed by the medical staff of department of critical care medicine of the Eighth Medical Center of People's Liberation Army General Hospital and department of critical care medicine of Beijing Friendship Hospital of Capital Medical University, and was obtained the National Utility Model Patent of China (ZL 2020 2 1476518.X). The device consists of the main pipe, first pipe sleeve, second pipe sleeve, valve body, joystick, axial sleeve, etc., which can not only effectively avoid air pollution and reduce the risk of cross-infection, but also reduce noise pollution and the negative impact of loud noise on patients and medical staff.

Design and application of flushing device of a totally enclosed sputum suction tube for reducing the incidence of ventilator-associated pneumonia / 中华危重病急救医学

Ventilator-associated pneumonia (VAP) is one of the most common infectious diseases in patients undergoing mechanical ventilation in intensive care unit (ICU). Although the use of closed sputum suction tube for sputum suction is the most common nursing measure for patients with tracheal intubation in ICU, and it is also an important measure to reduce the occurrence of VAP, the existing clinical technology still needs to use external flushing solution for humidification and flushing. In the process of use, the flushing operation has the risk that the amount of flushing fluid cannot be controlled, resulting in a large amount of flushing fluid entering the patient's airway, causing the patient to suffocate. The sputum attached to the wall of the sputum suction tube cannot be completely flushed, and the prolonged retention of the sputum leads to the proliferation of colonized bacteria, aggravating the infection of patients. Repeated pipe flushing not only increases consumables, but also increases the workload of medical staff. For this reason, the author has designed a fully enclosed sputum suction tube flushing device, which has obtained the National Utility Model Patent of China (patent number: CN 2019 2 1198740.5). This device is an integrated design, and the materials used for sputum suction, humidification and flushing are fully enclosed, without air exposure, and can be operated strictly without bacteria. Pulse flushing with syringe can clean the sputum attached to the inner wall and reduce the incidence of VAP. The volume of flushing fluid can be effectively controlled during use to ensure patient safety and reduce waste. It is worth popularizing and applying in clinical practice to optimize the operation steps and reduce the workload of medical staff.

Monitoring non-small cell lung cancer progression and treatment response through hyaluronic acid in sputum

We evaluated whether hyaluronan (HA) levels in the sputum could be used as a noninvasive tool to predict progressive disease and treatment response, as detected in a computed tomography scan in non-small cell lung cancer (NSCLC) patients. Sputum samples were collected from 84 patients with histological confirmation of NSCLC, 33 of which were in early-stage and 51 in advanced-stage disease. Patients received systemic chemotherapy (CT) after surgery (n=36), combined CT and immunotherapy (IO) (n=15), or targeted therapy for driver mutation and disease relapse (N=4). The primary end-point was to compare sputum HA levels in two different concentrations of hypertonic saline solution with overall survival (OS) and the secondary and exploratory end-points were radiologic responses to treatment and patient outcome. Higher concentrations of HA in the sputum were significantly associated to factors related to tumor stage, phenotype, response to treatment, and outcome. In the early stage, patients with lower sputum HA levels before treatment achieved a complete tumor response after systemic CT with better progression-free survival (PFS) than those with high HA levels. We also examined the importance of the sputum HA concentration and tumor response in the 51 patients who developed metastatic disease and received CT+IO. Patients with low levels of sputum HA showed a complete tumor response in the computed tomography scan and stable disease after CT+IO treatment, as well as a better PFS than those receiving CT alone. HA levels in sputum of NSCLC patients may serve as a candidate biomarker to detect progressive disease and monitor treatment response in computed tomography scans.

Educational technologies to guide pulmonary tuberculosis sputum collection: a systematic review / Tecnologías educativas para orientación de colecta de esputo de la tuberculosis pulmonar: revisión sistemática / Tecnologias educativas para orientação da coleta de escarro da tuberculose pulmonar: revisão sistemática

ABSTRACT Objective: To evaluate the contributions of educational technologies used during the guidelines for sputum collection from pulmonary tuberculosis. Method: Systematic review guided by Preferred Reporting items for Systematic Reviews with protocol registered in the database International Prospective Register of Systematic Reviews, with number CRD42020208162. The search was performed in the Cinahl, Scopus, PubMed, Web of Science, Embase, Lilacs, CENTRAL, CAPES, Proquest, OpenGrey databases and manual search in the reference list. The search, selection of studies, data extraction, and methodological evaluation using the Cochrane Risk-of-Bias tool were performed by two independent reviewers. Results: A total of 2,488 studies were evidenced, with seven being selected and analyzed, of which four used structured lectures; three, educational booklet; and one, an educational video, used alone or together, impacting the bacteriological confirmation of tuberculosis. The studies had a low risk of bias. Conclusion: Scientific evidence has shown that educational technologies contribute to increasing the quality, volume, and appearance of the sputum sample, which improves the bacteriological confirmation of the disease.

RESUMEN: Objetivo: Evaluar las contribuciones de las tecnologías educativas utilizadas durante las orientaciones para colecta de esputo de la tuberculosis pulmonar. Método: Revisión sistemática guiada por el Preferred Reporting items for Systematic Reviews con protocolo registrado en la base International Prospective Register of Systematic Reviews, bajo el número CRD42020208162. La búsqueda fue realizada en las bases de datos Cinahl, Scopus, PubMed, Web of Science, Embase, Lilacs, CENTRAL, CAPES, Proquest, OpenGrey y búsqueda manual en la lista de referencias. La búsqueda, selección de los estudios, extracción de los datos y evaluación metodológica por la herramienta del Riesgo de Sesgo Cochrane fueron realizadas por dos revisores independientes. Resultados: Se evidenciaron 2488 estudios, siendo siete seleccionados y analizados, de los cuales cuatro utilizaron palestras estructuradas, tres folder educativo y un vídeo educativo, utilizados de forma aislada o conjunta, impactando en la confirmación bacteriológica de la tuberculosis. Los estudios presentaron bajo riesgo de sesgo. Conclusión: Las evidencias científicas demostraron que las tecnologías educativas presentan como contribuciones el aumento de la calidad, volumen y aspecto de la muestra de esputo, lo que proporciona el perfeccionamiento de la confirmación bacteriológica de la enfermedad.

RESUMO Objetivo: Avaliar as contribuições das tecnologias educativas utilizadas durante as orientações para coleta de escarro da tuberculose pulmonar. Método: Revisão sistemática guiada pelo Preferred Reporting items for Systematic Reviews com protocolo registrado na base International Prospective Register of Systematic Reviews, sob o número CRD42020208162. A busca foi realizada nas bases de dados Cinahl, Scopus, PubMed, Web of Science, Embase, Lilacs, CENTRAL, CAPES, Proquest, OpenGrey e busca manual na lista de referências. A busca, seleção dos estudos, extração dos dados e avaliação metodológica pela ferramenta do Risco de Viés da Cochrane foram realizadas por dois revisores independentes. Resultados: Evidenciaram-se 2488 estudos, sendo sete selecionados e analisados, dos quais quatro utilizaram palestras estruturadas; três, folder educativo; e um, vídeo educativo. Todos foram utilizados de forma isolada ou em conjunto, impactando na confirmação bacteriológica da tuberculose. Os estudos apresentaram baixo risco de viés. Conclusão: As evidências científicas demonstraram que as tecnologias educativas apresentam como contribuições o aumento da qualidade, volume e aspecto da amostra de escarro, o que proporciona a melhoria da confirmação bacteriológica da doença.

Effects of different sputum suction pressure on intracranial pressure of severe craniocerebral injury patients / 中国实用护理杂志

Objective:To explore the best negative pressure standard of sputum aspiration in patients with severe craniocerebral injury, so as to achieve the best sputum aspiration effect and maintain relatively stable intracranial pressure.Methods:Totally 120 patients with severe craniocerebral injury admitted to the Department of Neurosurgery of Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from January to December 2019 were selected by convenience sampling method. According to the random number table method, patients were divided into 4 different negative pressure sputum aspiration groups: 75 mmHg (1 mmHg=0.133 kPa) group, 100 mmHg group, 150 mmHg group, 200 mmHg group, with 30 patients in each group. The levels of intracranial pressure and SpO 2, sputum cleaning time and mucosal injury were observed and compared in 4 groups: 1 min before sputum aspiration, 1 min after sputum aspiration and during sputum aspiration. Results:Comparison of intracranial pressure and SpO 2 in sputum aspiration among the four groups: intracranial pressure in 200 mmHg group was (22.23 ± 4.80) mmHg, which was higher than 75 mmHg group (16.33 ± 2.71) mmHg, 100 mmHg group (17.70 ± 2.32) mmHg, 150 mmHg group (17.20 ± 2.11) mmHg ( q=8.29, 6.58, 7.23), and SpO 2 was 0.906 ± 0.048, which was lower than 0.956 ± 0.013, 0.946 ± 0.018, 0.952 ± 0.023 in the other three groups ( q=7.81, 6.08, 6.69), with statistical significance (all P<0.01). There were statistically significant differences in intracranial pressure ( F=10.55) and SpO 2 ( F=36.57) among the four groups 1 min after sputum aspiration ( P<0.01);the 200 mmHg group had the highest intracranial pressure (15.63 ± 3.64) mmHg; the 100 mmHg group was (12.50 ± 1.48) mmHg and 150 mmHg group was (12.60 ± 1.00) mmHg, which had lower intracranial pressure than the 75 mmHg group (14.03 ± 2.86) mmHg. The sputum cleaning time was (35.53 ± 5.71) s in 75 mmHg group which was the longest and the shortest in 200 mmHg group (24.27 ± 3.22) s, with statistical significance among the four groups ( F=47.81, P<0.01). The incidence of airway mucosal injury in 200 mmHg group was 33.33% (10/30),which was higher than that in the other three groups, and the difference was statistically significant ( χ2=15.41, P<0.01). Conclusions:For patients with severe craniocerebral injury, artificial airway sputum viscosityⅠ-Ⅱ degree, suction negative pressure of 100-150 mmHg can better maintain the stability of intracranial pressure, control the decline of SpO 2 and prevent the damage of airway mucosa, and effectively reduce the adverse effects of sputum aspiration on patients with severe craniocerebral injury.

Design and application of a new heat-and-moisture exchanger with anti-splash sputum suctioning function / 中华危重病急救医学

In the emergency department, open endotracheal suctioning for mechanically ventilated patients with endotracheal intubation will lead to the spread of respiratory droplets and aerosols, polluting the surrounding environment and medical staff. The traditional heat-and-moisture exchanger has the effect of warming and humidifying, and can block pathogenic microorganisms, but it does not have the function of inserting a sputum suction tube. When the heat-and-moisture exchanger is pulled out for sputum suction, it is easy to cause sputum splash, which pollutes the surrounding environment and medical personnel. The addition of closed sputum suction devices will increase the economic burden on patients. Thus, the medical staff of emergency department of the First People's Hospital of Tongxiang City of Zhejiang Province designed a new type of heat-and-moisture exchanger with anti-splash sputum suctioning function and obtained the National Utility Model Patent of China (ZL 2021 2 0017615.0). The new heat-and-moisture exchanger is mainly composed of a receiving cavity, a connecting tube, a sputum suction tube intubation tube, a sealing valve, etc. The disposable sputum suction tube can be used to insert sputum suction, and at the same time, it can prevent the secretion from splashing to ensure sealing. The patent combines the humidification and pathogen blocking functions of the heat-and-moisture exchanger with the anti-splash sputum suctioning function, which is suitable for use in the emergency and critical care medicine departments and has clinically practical value.

Evaluación de spoligotyping a partir de baciloscopías como metodología alterna e independiente de cultivo para la genotipificación de Mycobacterium tuberculosis / Evaluation of spoligotyping-smear sputum as an alternative and culture-independent methodology for Mycobacterium tuberculosis genotyping

INTRODUCCIÓN: Tuberculosis (TBC) sigue siendo la segunda causa de muerte por una enfermedad infecto-contagiosa después del síndrome de inmunodeficiencia humana adquirida (SIDA). Actualmente, el escenario de técnicas y metodologías de laboratorio para la identificación y drogo-sensibilidad está cambiando gradualmente. Se han recomendado e introducido ensayos rápidos basados en la amplificación de ácidos nucleicos (NAAT's) que se desarrollan mediante la reacción de polimerasa en cadena (RPC). Bajo este principio, se destaca spoligotyping -una herramienta de genotipificación y epidemiología molecular en TBC- estandarizada a partir de aislados bacterianos, que permite el estudio del genoma de Mycobacterium mediante la amplificación de 43 secuencias cortas no repetitivas, localizadas en la región de repetición directa (RD1). OBJETIVO: Evaluación de spoligotyping a partir de baciloscopías, como una opción independiente de cultivo, para la caracterización de Mycobacterium tuberculosis a partir de muestras de esputo en pacientes del Instituto Nacional Cardiopulmonar de Tegucigalpa, Honduras. MÉTODO: De 37 pacientes con cultivo (y baciloscopía) positivos para M. tuberculosis, se obtuvieron 50 muestras de expectoración. Se realizó estudio microbiológico y molecular en muestras respiratorias conteniendo ADN de micobacterias, a partir de baciloscopías, concentrados y cultivo, para la identificación y análisis genotípico a través de la técnica de spoligotyping. RESULTADOS: El spoligotyping fue positivo en 37/37 de muestras de cultivo positivo (S: 100%), en 36/37 (S: 97,3%) de muestras con baciloscopía positiva y en 6/10 (S: 60%) de muestras de concentrado de esputo. La intensidad de la baciloscopía positiva tuvo una relación directa con la sensibilidad de spoligotyping. DISCUSIÓN: El fusionar el potencial de una herramienta útil en epidemiología molecular para analizar muestras de ADN proveniente de baciloscopías, visualiza una plataforma diagnóstica y genotípica para países en vías de desarrollo como una alternativa innovadora y altamente sensible en la hibridación de oligonucleótidos específicos a partir material genético en baciloscopías (P+, P++, P+++), pero requiere mejorar la concordancia entre patrones genéticos obtenidos, comparables con el uso estandarizado de aislados de cepas de M. tuberculosis.

BACKGROUND: Tuberculosis (TB) remains as the second cause of death by an infectious disease preceded by the acquired immune deficiency syndrome (AIDS). Currently, laboratory techniques and methodologies of diagnosis and drug susceptibility testing are constantly changing. Therefore, it has been recommended the introduction of rapid assays based on the amplification of nucleic acids test (NAAT's) through a polymerase chain reaction (PCR). Based on this principle, outstands spoligotyping - as a genotype and molecular epidemiology tool in tuberculosis - it is standardized to use isolated bacteria for the study of Mycobacterium genome through the amplification of 43 non-repetitive sequences, located at the direct repetitive region 1 (RD1). AIM Evaluation of spoligotyping from acid fast staining smears as an independent option from bacterial isolation to characterize Mycobacterium tuberculosis by using sputum samples from TB patients from National Cardiopulmonary Institute in Tegucigalpa, Honduras. METHOD: Of 37 patients with positive culture (and smear microscopy) for M. tuberculosis, 50 expectoration samples were obtained. Microbiological and molecular tests were performed in respiratory samples containing mycobacterial DNA from sputum smears, concentrates and solid culture, for identification and genotype analysis by spoligotyping technique. RESULTS: Spoligotyping was positive in 37/37 of positive culture samples (S: 100%), in 36/37 (S: 97.3%) of smear-positive samples and in 6/10 (S: 60%) of concentrate samples sputum. The intensity of positive smear microscopy had a direct relationship with the sensitivity of spoligotyping. DISCUSSION: This study combined the potential of a molecular epidemiology tool to analyse DNA from sputum samples in smears acid fast staining, it visualizes diagnosis and genotyping platform in developing countries gathering innovation and high sensitivity in the hibridization of specific olignonucleotides from positive smears (P+, P++, P+++). However, the low specificity showed the need to improve better agreement among genetic patterns compared to the standardized bacterial isolation from M. tuberculosis strains.

Caracterización de los pacientes con tuberculosis ingresados en Hospital Regional de Malabo / Characterization of the patients with tuberculosis admitted in the Regional Hospital of Malabo

RESUMEN Introducción: hace más de un siglo, Robert Koch identificó el microorganismo que causa la tuberculosis en el hombre, el Mycobacterium tuberculosis. Sin embargo, aún constituye un grave problema epidemiológico a nivel mundial. Se presentó un estudio realizado en los pacientes ingresados en el Servicio de Infeccioso del Hospital Regional de Malabo. Objetivo: caracterizar los pacientes con tuberculosis ingresados en la primera fase de tratamiento antituberculoso. Materiales y métodos: se realizó un estudio descriptivo, retrospectivo de los pacientes ingresados en el Servicio de Tratamiento de Infecciosos del Hospital Regional de Malabo. Periodo correspondiente a los meses de enero a julio del 2016. El universo fue 172 pacientes con diagnóstico de tuberculosis en todas sus formas clínicas. Los datos se obtuvieron en las historias clínicas. Resultados: predominó el sexo masculino en el 57,5 % de los pacientes y el grupo etáreo de 25 a 34 años con el 33,7 %. La baciloscopia negativa representó el 67,9 % de los casos y el estado nutricional más frecuente fue el bajo peso con 50,09 %. La tuberculosis en su forma pulmonar predominó en el 95,9 % de los pacientes. El 65,6 % de los pacientes fueron VIH negativos. La mejoría clínico radiológica al terminar el tratamiento fue de un 87,7 %. Conclusiones: el 57,5 % de los pacientes fueron masculinos entre 25 y 34 años de edad. Predominó la Baciloscopia negativa y la tuberculosis en su forma pulmonar. La mejoría clínica radiológica fue superior al terminar el tratamiento (AU).

ABSTRACT Introduction: More than a century ago, Robert Koch identified the microorganism causing tuberculosis in people: Mycobacterium tuberculosis. But it is still a serious epidemiologic problem around the world. The authors presented a study carried out in patients admitted in the Infectious Service of the Regional Hospital of Malabo. Objective: to characterize the patients with tuberculosis admitted in the first stage of the anti-tuberculosis treatment. Materials and methods: a descriptive, retrospective study was carried out on the patients who were admitted in the Infectious Service of the Regional Hospital of Malabo in the period from January to July 2016. The universe were 172 patients with diagnosis of tuberculosis in all its clinical forms. Data were gathered from the clinical records. Results: male sex predominated in 57.5 % of the patients as it also did the 25-34 years-old age group with 33.7 %. Negative sputum smear represented 67.9 % of the cases and the most frequent nutritional status was the low weight with 50.9 %. Tuberculosis in its pulmonary form predominated in 95.9 % of the patients. 65.6 % of the patients were HIV negative. The clinical radiological improvement at the end of the treatment was 87.7 %. Conclusions: 57.5 % of the patients were male aged 25-34 years old. Negative sputum smear and the pulmonary form of tuberculosis predominated. The clinical radiological improvement was higher at the end of the treatment (AU).

The yield of Acid-Fast Bacilli (AFB) and tuberculosis (TB) culture in the microbiologic diagnosis of childhood tuberculosis using sputum induction: A randomized controlled trial with interrupted time series

@#<p style="text-align: justify;"><strong>Objectives:</strong> This study aims to determine the diagnostic yield and safety of sputum induction with hypertonic saline in the microbiologic confirmation of childhood tuberculosis (TB) in a tertiary hospital in the Philippines.</p><p style="text-align: justify;"><strong>Methods:</strong> This is a randomized controlled trial with an interrupted time series in the control group. One hundred twelve (112) pediatric patients (4-18 years old) with clinical findings suggestive of TB were enrolled in the study. Patients were randomized into two groups composed of 56 patients each. Group A patients underwent sputum induction. Group B patients underwent spontaneous expectoration followed by sputum induction. The microbiologic yield for acid-fast bacilli and TB culture were determined and analyzed.</p><p style="text-align: justify;"><strong>Results:</strong> Among the patients randomized to Group A, microbiologic confirmation for TB was 8/56 patients (14.3%) after sputum induction. For patients randomized to Group B, microbiologic yield was 4/56 patients (7.1%) from spontaneous expectoration; after sputum induction, the microbiologic yield increased to 5/56 patients (8.9%). There is insufficient evidence of statistical significance in microbiologic yield on parallel analysis of the two separate groups (p=0.22). Furthermore, for patients randomized to Group B, the increase in microbiologic yield after sputum induction compared to spontaneous expectoration did not reach statistical significance (p=1.000). The procedure was well-tolerated among children; no serious adverse events were observed.</p><p style="text-align: justify;"><strong>Conclusion:</strong> Sputum induction is a feasible and safe method of specimen collection for microbiologic diagnosis of TB among children. While the microbiologic yield increased after sputum induction compared to spontaneous expectoration, the additional yield does not seem to be significant.</p>

Tuberculosis miliar / Miliary tuberculosis

Introducción: La tuberculosis es considerada como la enfermedad infecciosa más importante del mundo, a pesar de los esfuerzos que se han invertido para su control. Es producida por el complejo mycobacterium tuberculosis. El órgano más afectado es el pulmón, aunque puede tener repercusión extrapulmonar. Objetivo: Presentar el caso de un paciente diagnosticado de tuberculosis miliar a través del esputo posbroncoscopia. Caso clínico: Se expone el caso de un paciente diagnosticado de una tuberculosis miliar al cual se le realizó broncoscopia diagnóstica y lavado bronquial para bacilo ácido alcohol resistente (BAAR), el cual fue negativo. El diagnóstico se obtuvo por esputo BAAR posbroncoscopia. Conclusiones: El esputo posbroncoscopia es una opción con adecuada rentabilidad en el diagnóstico de esta enfermedad infecciosa(AU)

Introduction: Tuberculosis is considered the most important infectious disease in the world, despite the efforts that have been invested to control it. It is produced by the mycobacterium tuberculosis complex. The most affected organs are the lungs, although it can have extrapulmonary repercussions. Objective: To report the case of a patient diagnosed with miliary tuberculosis through post-bronchoscopy sputum. Clinical case report: The case of a patient diagnosed with miliary tuberculosis is reported. This patient underwent diagnostic bronchoscopy and bronchial lavage for acid-alcohol-resistant bacillus (ABB), which was negative. The diagnosis was obtained by postbronchoscopy ARB sputum. Conclusions: Post-bronchoscopy sputum is an option, with adequate profitability in the diagnosis of this infectious disease(AU)

Isolation of and risk factors for airway infection with Pseudomonas aeruginosa in patients with non-cystic fibrosis bronchiectasis / Isolamento de Pseudomonas aeruginosa e fatores de risco de infecção das vias aéreas pela bactéria em pacientes com bronquiectasias não fibrocísticas

ABSTRACT Objective: To identify microorganisms in sputum samples of patients with stable non-cystic fibrosis bronchiectasis and to determine risk factors related to the isolation of Pseudomonas aeruginosa (PA) in those patients. Methods: Consecutive patients were recruited from a tertiary hospital outpatient clinic in the city of Fortaleza, Brazil. The patients were submitted to spirometry, six-minute walk test, HRCT, and sputum collection. Data on serum fibrinogen levels, disease severity, sputum color, and history of azithromycin treatment were collected. Results: The study included 112 patients, and females predominated (68%). The mean age was 51.6 ± 17.4 years. Most patients presented with mild-to-moderate disease (83%). The mean six-minute walk distance was 468.8 ± 87.9 m. Mean FEV1 and FVC, in % of predicted values, were 60.4 ± 21.8% and 69.9 ± 18.5%, respectively. The mean serum fibrinogen level was 396.1 ± 76.3 mg/dL. PA was isolated in 47 patients, other potentially pathogenic microorganisms (PPMs) were isolated in 31 patients, and non-PPMs were isolated in 34 patients. Purulent sputum was identified in 77 patients (68%). The patients with PA, when compared with those without it, presented with more severe disease, higher serum fibrinogen levels, and lower FVC%. In addition, purulent sputum and long-term azithromycin treatment were more common in those with PA. The multivariate regression analysis showed that the independent factors associated with PA were serum fibrinogen level > 400 mg/dL (OR = 3.0; 95% CI: 1.1-7.7) and purulent sputum (OR = 4.3; 95% CI: 1.6-11.3). Conclusions: In our sample, the prevalence of PA in sputum was 42%. Sputum color and inflammatory markers were able to predict the isolation of PA, emphasizing the importance of routine sputum monitoring.

RESUMO Objetivo: Identificar microrganismos em amostras de escarro de pacientes com bronquiectasia não fibrocística estável e determinar os fatores de risco relacionados com o isolamento de Pseudomonas aeruginosa (PA) nesses pacientes. Métodos: Pacientes consecutivos foram recrutados em um ambulatório de um hospital terciário em Fortaleza (CE). Os pacientes foram submetidos a espirometria, teste de caminhada de seis minutos, TCAR e coleta de escarro. Foram coletados dados referentes ao fibrinogênio sérico, gravidade da doença, cor do escarro e histórico de tratamento com azitromicina. Resultados: O estudo incluiu 112 pacientes, com predomínio do sexo feminino (68%). A média de idade foi de 51,6 ± 17,4 anos. A maioria dos pacientes apresentou doença leve a moderada (83%). A média da distância percorrida no teste de caminhada de seis minutos foi de 468,8 ± 87,9 m. A média do VEF1 em % do previsto foi de 60,4 ± 21,8%, e a da CVF em % do previsto foi de 69,9 ± 18,5%. A média do fibrinogênio sérico foi de 396,1 ± 76,3 mg/dL. PA foi isolada em 47 pacientes; outros microrganismos potencialmente patogênicos (MPP) foram isolados em 31; não MPP foram isolados em 34. Escarro purulento foi identificado em 77 pacientes (68%). Os pacientes com PA, em comparação com aqueles sem, apresentaram doença mais grave, fibrinogênio sérico mais elevado e menor CVF%. Além disso, escarro purulento e tratamento prolongado com azitromicina foram mais comuns naqueles com PA. A análise de regressão multivariada mostrou que os fatores independentes relacionados com PA foram fibrinogênio sérico > 400 mg/dL (OR = 3,0; IC95%: 1,1-7,7) e escarro purulento (OR = 4,3; IC95%: 1,6-11,3). Conclusões: Em nossa amostra, a prevalência de PA no escarro foi de 42%. A cor do escarro e os marcadores inflamatórios foram capazes de prever o isolamento de PA, o que enfatiza a importância do monitoramento rotineiro do escarro.

Bleach-concentrated direct sputum smear microscopy procedure for diagnosis of pulmonary tuberculosis in poverty areas / Microscopia direta de amostra de escarro concentrado em água sanitária para diagnóstico de tuberculose pulmonar

ABSTRACT INTRODUCTION: Pulmonary tuberculosis caused by Mycobacterium tuberculosis is a serious public health problem affecting millions of people worldwide. The development of easy and low-cost diagnostic methods is crucial for disease control in rural remote and poverty areas and among vulnerable groups. OBJECTIVE: To evaluate the accuracy of laboratory methods for the diagnosis of Pulmonary tuberculosis. MATERIAL AND METHODS: Sputum samples from patients with clinical signs and symptoms were analyzed by microscopy after chemical treatment and spontaneous sedimentation and compared with methods employed routinely: direct sputum smear microscopy, culture, and GeneXpert®MTB/RIF. RESULTS: From the samples analyzed, 16% were positive by microscopy in the processed samples, 18% by both culture and Xpert®MTB/RIF, while 13% in the direct microscopy. The processed samples showed a 31% increase in positivity (57 samples) compared to conventional direct microscopy. In the analysis of the accuracy of the evaluated methods, all the results were statistically significant proving that they were not randomly positive or negative and confirming that there is a tendency for these results. CONCLUSION: Chemical treatment and spontaneous sedimentation of the sputum samples procedure represent an effective diagnostic tool in situations where more advanced technologies are not feasible. Besides the higher accuracy and greater detection of positive cases regarding the direct smear, the procedure strengthens biosafety by decreasing the risks of aerial contamination by Mycobacterium tuberculosis for laboratory professionals.

RESUMEN INTRODUCCIÓN: La tuberculosis pulmonar causada por Mycobacterium tuberculosis es un grave problema de salud pública que afecta millones de individuos en el mundo. El desarrollo de métodos de diagnóstico fáciles y de bajo costo es esencial para el control de la enfermedad en zonas rurales remotas y pobres y entre los grupos vulnerables. OBJETIVO: Evaluar la exactitud de métodos de laboratorio para el diagnóstico de tuberculosis pulmonar. MATERIAL Y MÉTODO: Las muestras del esputo de pacientes con signos y síntomas clínicos fueron analizadas por microscopía luego de tratamiento químico y sedimentación espontánea y comparados con métodos empleados ordinariamente: baciloscopía directa de esputo, cultivo y GeneXpert® MTB/RIF. RESULTADOS: Entre las muestras analizadas, 16% fueron positivas por microscopía en las muestras procesadas; 18% por cultivo y Xpert® MTB/RIF; y 13% por microscopía directa. Las muestras procesadas presentaran un aumento de 31% de positividad (57 muestras) con respecto a la microscopía directa convencional. En el análisis de los métodos, todos los resultados fueron estadísticamente significativos, comprobando que no eran aleatoriamente positivos o negativos y confirmando que hay una tendencia para esos resultados. CONCLUSIÓN: El tratamiento químico y la sedimentación espontánea de las muestras de esputo representan una herramienta diagnóstica eficaz en las situaciones en las cuales tecnologías más avanzadas no son viables. Además de la mayor precisión y mayor detección de casos positivos de lo que hace el frotis directo, el procedimiento fortalece la bioseguridad, disminuyendo los riesgos de contaminación del aire por Mycobacterium tuberculosis para el personal de laboratorio.

RESUMO INTRODUÇÃO: A tuberculose pulmonar causada por Mycobacterium tuberculosis é um grave problema de saúde pública que afeta mundialmente milhões de indivíduos. O desenvolvimento de métodos de diagnóstico fáceis e de baixo custo é essencial para o controle da doença nas áreas rurais remotas e de pobreza e entre os grupos vulneráveis. OBJETIVO: Avaliar a acurácia dos métodos laboratoriais para o diagnóstico de tuberculose pulmonar. MATERIAL E MÉTODOS: As amostras de escarro de pacientes com sinais e sintomas clínicos foram analisadas por microscopia após tratamento químico e sedimentação espontânea e comparadas com métodos empregados rotineiramente: baciloscopia direta do escarro, cultura e GeneXpert® MTB/RIF. RESULTADOS: Das amostras analisadas, 16% foram positivas por microscopia nas amostras processadas; 18%, por cultura e Xpert® MTB/RIF; e 13%, por microscopia direta. As amostras processadas apresentaram um aumento de 31% de positividade (57 amostras) em relação à microscopia direta convencional. Na análise dos métodos avaliados, todos os resultados foram estatisticamente significativos, comprovando que não eram positivos ou negativos aleatoriamente e confirmando que há uma tendência para esses resultados. CONCLUSÃO: O tratamento químico e a sedimentação espontânea das amostras de escarro representam uma ferramenta diagnóstica eficaz nas situações em que tecnologias mais avançadas não são viáveis. Além da maior precisão e maior detecção de casos positivos em relação ao esfregaço direto, o procedimento reforça a biossegurança, diminuindo os riscos de contaminação aérea por Mycobacterium tuberculosis para profissionais de laboratório

Effects of lung rehabilitation training combined with systematic expectoration training on rehabilitation of stroke in patients subjected to tracheotomy / 中国基层医药

Objective:To investigate the effects of lung rehabilitation training combined with systematic expectoration training on rehabilitation of stroke in patients subjected to tracheotomy.Methods:Sixty patients with stroke subjected to tracheotomy who received treatment in Ningbo Rehabilitation Hospital, China between January 2018 and July 2019 were included in this study. These patients were randomly assigned to undergo either systematic expectoration training alone (control group, n = 30) or lung rehabilitation training combined with systematic expectoration training (observation group, n = 30) for 1 month. Therapeutic effect, number of daily sputum aspirations, blood oxygen saturation, maximum inspiratory volume, maximum expiratory volume, duration of infection control, duration of placement of indwelling tracheal catheter, and complications were compared between the two groups. Results:Total effective rate in the observation group was significantly higher than that in the control group [90.0% (27/30) vs. 73.3% (22/30), χ2 = 5.092, P < 0.05]. Before treatment, there were no significant differences in the number of daily sputum aspirations, blood oxygen saturation, maximum inspiratory volume, and maximum expiratory volume between the two groups ( t = 1.139, 0.705, 0.253, 0.519, P > 0.05). After treatment, the number of daily sputum aspirations in the observation group was significantly lower than that in the control group [(2.16 ± 1.28) counts vs. (3.59 ± 2.39) counts, t = 5.616]. Blood oxygen saturation, maximum inspiratory volume, maximum expiratory volume in the observation group were (99.3 ± 0.5)%, (479.37 ± 64.15) mL and (482.23 ± 62.16) mL, respectively, which were significantly higher than those in the control group [(95.8 ± 0.7)%, (403.58 ± 62.13) mL, (398.37 ± 64.24) mL, t = 4.075, 4.279, 4.106, all P < 0.05]. The duration of infection control and the duration of placement of the indwelling tracheal catheter in the observation group were (15.21 ± 5.48) days and (23.39 ± 6.51) days, which were significantly shorter than those in the control group [(23.39 ± 6.51) days, (32.64 ± 6.16) days, t = 6.080, 5.119, both P < 0.05]. The incidences of pulmonary infection and gastroesophageal reflux in the observation group were 6.7% (2/30) and 6.7% (2/30), respectively, which were significantly lower than those in the control group [16.7% (5/30), 70.0% (21/30), χ2 = 4.071, 4.372, both P < 0.05]. The proportion of patients with respiratory muscle strength ≥ grade 4 in the observation group was significantly higher than that in control group [96.7% (29/30) vs. 70.0% (21/30), χ2 = 14.402, P < 0.05]. Conclusion:Lung rehabilitation training combined with systematic expectoration training can greatly promote the rehabilitation of stroke patients subjected to tracheotomy, improve respiratory function, reduce pulmonary infection, and is safe and reliable.

The clinical value of induced sputum DKK3 gene methylation in the evaluation of disease condition and prognosis for non-small cell lung cancer / 中国医师进修杂志

Objective:To study the clinical value of induced sputum DKK3 gene methylation in the evaluation of disease condition and prognosis for non-small cell lung cancer (NSCLC).Methods:Eighty NSCLC patients (observation group) and 50 benign lung disease patients (control group) who were treated in Linshu County People′s Hospital of Shandong Province from January 2015 to December 2017 were enrolled. Methylation specific polymerase chain reaction (PCR) was used to detect the induced sputum DKK3 gene methylation. The DKK3 gene methylation rates in different clinicopathological factors were compared.Results:The DKK3 gene methylation rate in observation group was significantly higher than that in control group: 81.3%(65/80) vs. 2.0%(1/50), the difference was statistically significant ( P<0.05). The DKK3 gene was methylated in lung cancer cells, and was unmethylated in normal lung epithelial cells BEAS-2B. The DKK3 gene methylation rates had correlation with pleural effusion, degree of differentiation, tumor diameter, lymph node metastasis, distant metastasis and TNM staging ( P<0.05). The R0 radicalresection, 3-year survival rate and total survival time in patients with DKK3 gene methylated were significantly lower than those in patient with DKK3 gene unmethylated: 53.8%(35/65) vs. 15/15, 28.1% vs. 37.9%, (1.8 ± 0.3) years vs. (2.1 ± 0.6) years, the differences were statistically significant ( P<0.05). Conclusions:The induced sputum DKK3 gene methylation rate in NSCLC patients is significantly higher and is related with prognosis. The induced sputum DKK3 gene methylation may provide basis for evaluating of disease condition and prognosis for NSCLC patients.