[Summary of the clinical characteristics of tuberculous otitis media].

Objective:The purpose of this study was to analyze and summarize the clinical characteristics and diagnostic methods of tuberculous otitis media（TOM）, to enrich clinical experience in diagnosis and treatment of tuberculous otitis media, so as to reduce missed diagnosis and misdiagnosis, and facilitate timely and effective therapy for better prognosis. Methods:This study retrospectively analyzed the clinical data of patients with tuberculous otitis media who were hospitalized in the Ear ward of our hospital and received surgical treatment from 2008 to 2022. The data of patients' clinical characteristics, radiological examination, intraoperative findings and therapeutic strategies were recorded and summarized. Results:A total of 23 cases （26 ears） of tuberculous otitis media were included in this retrospective study. The most common clinical symptoms were otorrhea（thin odorless fluid）（100%） and conductive hearing loss（100%）, with a high incidence of facial paralysis（23.1%）. It was not sensitive to traditional antibiotic treatment, eg. Levofloxacin （50% effective rate only）, and relapsed soon after drug withdrawal. It was revealed that all the surgical views had gray and white tough granulation tissue hyperplasia（100%）, and 23.1% with caseous necrosis. The purpose of surgery was to clear the lesion, reduce the recurrence rate of suppurative infection, and repair the function （hearing reconstruction or facial nerve decompression） as appropriate. The paraffin pathology of granulation tissue were reported as typical granulomatous inflammation and caseous necrosis with positive acid-fast staining, which was consistent with tuberculosis. Conclusion:It was easily confused by the clinical manifestations of tuberculous otitis media and common chronic suppurative otitis media. When met with the following conditions, we should pay highly attention to suspect tuberculous otitis media: The severity of local manifestations did not match with the length of the disease; with poor tympanic membrane at the early stage with no obvious cholesteatomas, with facial paralysis or hearing loss early onset; insensitive to traditional antibiotic treatment; with extensive granulation appeared in the tympanum and or mastoid cavity, with or without caseous necrosis or dead bone in the early days. The diagnosis should be confirmed based on the acid-fast staining of the histopathological section to detect positive acid-fast bacilli. Meanwhile, multiple laboratory examination methods（such as T-spot and PCR） should be integrated synchronously to help support the diagnosis.

Nocardia pseudobrasiliensis infection in a patient with arthritis.

Nocardia pseudobrasiliensis is a new taxon constituting an emerging species of human pathogenic Nocardia, which shares morphological features with N. brasiliensis. However, N. pseudobrasiliensis is more invasive and more easily disseminated, and it exhibits distinctive antibiotic susceptibility. Few clinical cases related to N. pseudobrasiliensis infection have been reported, and N. pseudobrasiliensis hydrarthrosis has not been described. Here, we analyzed the case information, diagnostic process, treatment, and prognosis of a patient with N. pseudobrasiliensis hydrarthrosis who received treatment in Zhejiang Provincial People's Hospital. Magnetic resonance imaging showed joint cavity effusion and soft tissue swelling with high signal on proton density-fat saturated images and low signal on T1-weighted images. Oil microscopy revealed abundant acid-fast-positive filaments in hydrarthrosis puncture fluid. The pathogen was identified as N. pseudobrasiliensis by matrix-assisted laser desorption ionization-time of flight mass spectrometry. In contrast to the 100% ciprofloxacin resistance displayed by N. brasiliensis, this clinical isolate of N. pseudobrasiliensis was completely susceptible. In summary, this is the first report of N. pseudobrasiliensis in joint effusion from a patient with arthritis.

Pathomorphological characteristics of tuberculous placenta and its clinical implication.

BACKGROUND: The study of pathologic diagnosis of placental TB is rare. The aim of this study is analyzing the pathomorphological characteristics of tuberculosis (TB) placenta during pregnancy and its clinical significance. METHODS: Nineteen cases of placental tissue specimens during pregnancy were collected from June 2015 to February 2022 at Shanghai Public Health Clinical Center, the only inpatient center for pregnant women with TB in Shanghai, China. Hematoxylin-eosin staining, acid-fast staining, and molecular testing were applied to analyze them comprehensively in combination with clinical information. RESULTS: Among the 19 cases, 7 cases caused intrauterine stillbirth, 3 cases received artificial abortion required by the pregnant woman, the other 9 cases received standard delivery and the infants survived, however, 3 of them were low-weight preterm infants, and another 1 case suffered mild intrauterine asphyxia. The 9 surviving infants were followed-up, of which 3 cases got congenital TB. For pathological characteristics of placental tissues under light microscopy, there were 3 cases of epithelioid granuloma formation, 13 cases of acute fetal membranitis, 4 cases of caseous necrosis, 7 cases of inflammatory necrosis, 10 cases of coagulative necrosis, and 6 cases with small focal calcifications. All placental tissues were positive for acid-fast staining and polymerase chain reaction (PCR). Molecular pathological diagnosis showed that 18 cases were positive for Mycobacterium tuberculosis, with 1 case not having received examination. CONCLUSIONS: Combining acid-fast staining and molecular pathological testing is helpful for accurately diagnosing placental TB.

Subtyping of Cryptosporidium parvum Obtained from Humans and Calves in Van, Turkey.

Background: We aimed to investigate the prevalence of Cryptosporidium species detected in humans and calves in the Van region of Turkey. Methods: A total of 150 patients, comprising 60 who were immunosup-pressed, 50 who were immunosuppressed and had diarrhea, and 40 who had only diarrhea, were enrolled in this study in the Department of Medical Parasitology, Van Yuzuncu Yil University Faculty of Medicine, Turkey. Stool samples were taken from the rectums of a total of 50 calves that had 30 diarrhea and 20 that did not have diarrhea, from the stables and farms of 10 central villages of Van, Turkey. All samples were analyzed using modified acid-fast staining, immunochromatographic test, and PCR. Cryptosporidium positive samples were also subtyped. Results: Only C. parvum subtypes were detected in all positive samples. C. parvum was detected in 30 (20%) of the 150 human stool samples, while it was detected in 5 (10%) of the 50 samples from the calves. The GP60 gene region was amplified and sent for sequence analysis to identify the C. parvum subtypes. Conclusion: As a result, C. parvum is found to be an active species that caused cryptosporidiosis is in the Van region. IIdA24G1 subtype of C. parvum were found in both human and calf. Therefore, due to the zoonotic feature of the C. parvum IIdA24G1 subtype, it has been shown that the calves in the region are a significant risk for humans.

Tuberculosis with cavities? Rapid diagnosis of Rhodococcus equi pulmonary infection with cavities by acid-fast staining: A case report.

Rhodococcus equi is a conditionally pathogenic bacterium widely distributed in soil, water, and marine environments, which can cause respiratory infections, pleurisy, blood and even bone marrow infections in immunocompromised people, and particularly in patients with acquired immunodeficiency syndrome (AIDS). This case report describes a patient with initially suspicion of tuberculosis (TB) as an outpatient in a TB clinic. However, laboratory findings identified R. equi in his sputum sample based on a positive acid-fast stain, which was highly suggestive of a pulmonary infection caused by R. equi. The patient was subsequently admitted to the respiratory unit for treatment. Once the source of infection was identified, the patient was treated with a combination of antibiotics for 2 weeks and was discharged with a significant improvement in symptoms.

Clinical performance of quantitative PCR for the molecular identification of skeletal tuberculosis from formalin-fixed paraffin-embedded tissues.

BACKGROUND: At present, skeletal tuberculosis (TB) diagnosis is mostly by histopathology, but the positivity rate is low. There is a need to develop new methods for the molecular identification of this disorder. Therefore, we aimed to investigate the clinical utility of quantitative PCR (qPCR)-based diagnosis of skeletal TB from formalin-fixed paraffin-embedded (FFPE) tissues and its comparative evaluation with acid-fast bacillus staining (AFS). METHODS: We detected Mycobacterium tuberculosis (M. tuberculosis/MTB) DNA using qPCR and AFS in FFPE tissue samples from 129 patients suspected of having skeletal TB. The sensitivity, specificity as well as area under the curve (AUC) of qPCR and AFS were calculated. Meanwhile, some factors potentially affecting qPCR and AFS results were investigated. RESULTS: Overall, qPCR outperformed AFS in detecting M. tuberculosis. The AUC of qPCR was higher than that of AFS (0.744 vs.0.561, p < 0.001). Furthermore, decalcification of bone tissues did not affect the sensitivity and specificity of qPCR tests. Whereas it impacted the performance of AFS, decalcification increased AFS's specificity and decreased its sensitivity (p < 0.05). Moreover, qPCR had a significantly larger AUC than AFS in decalcified and non-decalcified groups (0.735/0.756 vs. 0.582/0.534, p < 0.001) respectively. Similarly, the AUC of PCR was more extensive than that of AFS regardless of skeletal TB patients with concomitant pulmonary TB or not (0.929 vs. 0.762; 0.688 vs. 0.524, p < 0.01). CONCLUSIONS: Our data demonstrate that qPCR offers superior accuracy for the detection of mycobacteria in FFPE tissues compared to traditional AFS, indicating its clinical value in osteoarticular TB diagnosis.

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.

Application of acid-fast staining combined with GeneXpert MTB/RIF in the diagnosis of non-tuberculous mycobacteria pulmonary disease.

OBJECTIVE: To evaluate the clinical diagnostic value of positive acid-fast staining combined with negative GeneXpert MTB/RIF in the diagnosis of non-tuberculous mycobacteria pulmonary disease (NTM-PD). METHODS: A total of 133 inpatients with confirmed NTM-PD were included consecutively between January 1, 2018 and December 31, 2019, at Tongji Hospital and Jinyintan Hospital, Tongji Medical College of Huazhong University of Science and Technology, in Wuhan, China. One hundred patients with confirmed pulmonary tuberculosis (PTB) were randomly included as the control group. RESULTS: The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of positive acid-fast staining combined with a negative GeneXpert MTB/RIF result were 51.13% (95% confidence interval (CI) 42.52-59.73%), 97.00% (95% CI 93.60-100.40%), 95.78% (95% CI 90.98-100.57%), and 59.88% (95% CI 52.25-67.51%), respectively. When subjects were limited to patients with positive acid-fast staining, the sensitivity of a negative GeneXpert MTB/RIF result was 88.31% (95% CI 80.97-95.65%). When acid-fast staining was conducted ≥3 times, the sensitivity of this combination diagnosis method increased to 61.67% (95% CI 49.00-74.33%). CONCLUSIONS: Positive acid-fast staining combined with a negative GeneXpert MTB/RIF result could be an effective and time-saving method for the diagnosis of NTM-PD.

Comparative study on the diagnostic value of four pathogenic methods in the detection of Mycobacterium tuberculosis in sputum specimens / 公共卫生与预防医学

Objective To explore the different diagnostic values of acid-fast staining, tuberculosis culture, tuberculosis DNA detection (TB-DNA), tuberculosis RNA constant temperature amplification technology (SAT-TB) in the detection of Mycobacterium tuberculosis in sputum specimens. Methods A total of 200 pulmonary tuberculosis patients and 80 non-tuberculosis patients who were hospitalized in Hebei Chest Hospital between September 2015 and September 2019 were selected for this study. Sputum samples were collected after admission, and the detection values of acid-fast staining, tuberculosis culture, TB-DNA, and SAT-TB in sputum samples were statistically analyzed. Results The differences in the sensitivity, accuracy, positive predictive value, and negative predictive values of the four diagnostic methods of acid-fast staining, tuberculosis culture, TB-DNA, and SAT-TB were statistically significant (P TB-DNA> tuberculosis culture> acid-fast staining. In terms of the positive predictive value of diagnosis, the values of SAT-TB, TB-DNA, and tuberculosis culture were higher than that of acid-fast staining. The Kappa values of the four methods and the gold standard were: Kappa (acid-fast staining) = 0.145, Kappa (tuberculosis culture) = 0.395, Kappa (TB-DNA) = 0.602, and Kappa (SAT-TB) = 0.770. Conclusion The four diagnostic methods of acid-fast staining, tuberculosis culture, TB-DNA, and SAT-TB all had certain detection value with their advantages and disadvantages. SAT-TB was a better detection method with high specificity, good sensitivity, and a short detection timer, which could quickly identify bacteria and distinguish live bacteria.

Prevalence of Pulmonary Tuberculosis and Associated Factors Among HIV Positive Patients Attending Antiretroviral Therapy Clinic at Arba Minch General Hospital, Southern Ethiopia.

BACKGROUND: Tuberculosis (TB) is an extremely contagious disease detrimentally affecting virtually every organ, most importantly the lungs. Pulmonary complications have been one of the commonest causes of morbidity and mortality since the advent of AIDS (Acquired Immune Deficiency Syndrome) pandemic. The AIDS virus has considerably reshape the epidemiology of TB by widening the risk of reactivating latent TB, increasing the possibility of TB infection once contracted to tubercle bacilli (re-infection) and by elevating the risk of rapid progression instantly after the infection. In this background, this study is intended to understand the prevalence of pulmonary tuberculosis and associated factors amongst Human Immunodeficiency Virus (HIV) positive patients attending antiretroviral therapy (ART) clinic in Arba Minch General hospital during the study period (March to May, 2016). METHODS: A cross-sectional study was carried out at Arba Minch Hospital from March to May, 2016. To assess the associated factors, a pre-tested structured questionnaire has been used. Sputum samples were collected and examined microscopically by using acid fast staining. The data was analyzed using Statistical Package for Social Services, version 20. RESULTS: Totally, 291 HIV positive patients were included in this study of which 71.5% were females and 28.5% were males. It was found that 42.3% of respondents were in the age ranged between 31-40 years. Of the 291 patients screened, 21 were positively diagnosed with pulmonary TB making the overall prevalence rate of 7.2%. From this study, it was revealed that CD4 count, previous history of tuberculosis and smoking were the significant predictors of tuberculosis (p˂0.05) in HIV patients. CONCLUSION: The results of the present study envisaged that the prevalence of HIV/TB co-infection was 7.2%. Previous history of TB, CD4 count less than 200/µl, and smoking habit were the possible risk factors elucidated. Therefore, TB screening among HIV-positive patients, public awareness, and community mobilization should be encouraged.

Cytokinin Signaling in Mycobacterium tuberculosis.

It was recently reported that the human-exclusive pathogen Mycobacterium tuberculosis secretes cytokinins, which had only been known as plant hormones. While cytokinins are well-established, adenine-based signaling molecules in plants, they have never been shown to participate in signal transduction in other kingdoms of life. M. tuberculosis is not known to interact with plants. Therefore, we tested the hypothesis that cytokinins trigger transcriptional changes within this bacterial species. Here, we show cytokinins induced the strong expression of the M. tuberculosis gene Rv0077c. We found that Rv0077c expression is repressed by a TetR-like transcriptional repressor, Rv0078. Strikingly, cytokinin-induced expression of Rv0077c resulted in a loss of acid-fast staining of M. tuberculosis While acid-fast staining is thought to be associated with changes in the bacterial cell envelope and virulence, Rv0077c-induced loss of acid-fastness did not affect antibiotic susceptibility or attenuate bacterial growth in mice, consistent with an unaltered mycolic acid profile of Rv0077c-expressing cells. Collectively, these findings show cytokinins signal transcriptional changes that can affect M. tuberculosis acid-fastness and that cytokinin signaling is no longer limited to the kingdom Plantae.IMPORTANCE Cytokinins have only previously been known as plant hormones. The discovery that they can be used as signaling molecules outside of plants broadens the repertoire of small molecules that can potentially affect gene expression in all domains of life.

Mycobacteriosis and Tuberculosis: Laboratory Diagnosis.

BACKGROUND: Tuberculosis is one of the most important infectious diseases that has claimed its victims throughout much of known human history. With Koch's discovery of the tubercle bacillus as the etiologic agent of the disease, his sanitary and hygienic measures, which were based on his discovery and the development of a vaccine against tuberculosis by Albert Calmette and Camille Guérin in 1921, an attenuated Mycobacterium bovis strain, bacilli Calmette-Guérin (BCG), and the discovery of the first antibiotic against tuberculosis, streptomycin by Selman Waksman in 1943, soon led to the opinion that appropriate control measures had become available for tuberculosis and it had been assumed that the disease could ultimately be eradicated.The emergence of resistant strains of this bacteria and widespread distribution of the disease in the world, and the emergence of the AIDS epidemic destroyed any possibility of global control of tuberculosis in the foreseeable future. OBJECTIVES: The purpose of this review is to highlight the current scientific literature on mycobacterial infections and provide an overview on the laboratory diagnosis of tuberculosis and non-tuberculosis infections based on conventional phenotypic and modern molecular assays. METHOD: In this study, a number of 65 papers comprising 20 reviews, 9 case reports, and 36 original research in association with mycobacteriosis and the laboratory diagnosis of mycobacterial infections, were reviewed. RESULTS: Based on our analysis on the published documents methods applied for the laboratory diagnosis of tuberculosis are continually assessed and developed in order to achieve more rapid, less expensive, and accurate results. Acid-fast staining and culture for mycobacteria remain at the core of any diagnostic algorithm with the sensitivity of 20-70% and specificity of 95-98% for AFB microscopy and the sensitivity of 95% and the specificity of 98% for culture based diagnosis. Following growth in culture, molecular tests such as nucleic acid hybridization probes and DNA sequencing may be used for definitive species identification. Nucleic acid amplification methods provide the means for direct detection of Mycobacterium tuberculosis in respiratory specimens without the prerequisite to isolate or culture the organism, leading to more rapid diagnosis and better patient care. CONCLUSION: As the researchers in a developing country, we strongly believe that despite significant advances in laboratory capacity, in many countries reliable confirmation of suspected mycobacterial diseases is hindered by a lack of knowledge on proper standardized methods, sufficient funds, suitably trained staff and laboratory supplies.

[Cryptosporidium infection in Nanjing City from 2015 to 2016].

OBJECTIVE: To investigate the status of Cryptosporidium infection in the population in Nanjing City so as to provide the evidence for the prevention and control of cryptosporidiosis. METHODS: A total of 100 fecal samples were collected from each of three districts (Liuhe, Qixia and Gaochun) and one hospital (Nanjing Zhongda Hospital) in 2015 and 2016 respectively. The fecal samples were detected for Cryptosporidium with microscopy (by using the gold amine phenol-modified acid-fast staining) and the positive samples were detected again for the molecular biology confirming by using the fluorescence quantitative PCR. RESULTS: During the two years, 581 cases of normal population who lived in the city were surveyed and no Cryptosporidium infection was found. Among 202 cases of outpatients with chronic diarrhea, there were 9 Cryptosporidium positive cases with the microscope scanning method (4.46%), and among the 9 cases, 7 cases showed obvious logarithmic amplification curves showing positive Cryptosporidium nucleic acid, but 2 cases without the obvious logarithmic amplification curves, and the Cryptosporidium nucleic acid positive rate was 3.47%. CONCLUSIONS: Cryptosporidium infection is not found in the normal population of Nanjing City, but the Cryptosporidium infection is found in the chronic diarrhea patients. The results imply that we should strengthen the detection of Cryptosporidium in the chronic diarrhea patients, so as to provide the evidence for improving the diagnosis and treatment of cryptosporidiosis.

Cryptosporidium infection in Nanjing City from 2015 to 2016 / 中国血吸虫病防治杂志

Objective To investigate the status of Cryptosporidium infection in the population in Nanjing City so as to pro-vide the evidence for the prevention and control of cryptosporidiosis. Methods A total of 100 fecal samples were collected from each of three districts(Liuhe,Qixia and Gaochun)and one hospital(Nanjing Zhongda Hospital)in 2015 and 2016 respective-ly. The fecal samples were detected for Cryptosporidium with microscopy(by using the gold amine phenol-modified acid-fast staining)and the positive samples were detected again for the molecular biology confirming by using the fluorescence quantita-tive PCR. Results During the two years,581 cases of normal population who lived in the city were surveyed and no Cryptospo-ridium infection was found. Among 202 cases of outpatients with chronic diarrhea,there were 9 Cryptosporidium positive cases with the microscope scanning method (4.46％),and among the 9 cases,7 cases showed obvious logarithmic amplification curves showing positive Cryptosporidium nucleic acid,but 2 cases without the obvious logarithmic amplification curves,and the Cryptosporidium nucleic acid positive rate was 3.47％. Conclusions Cryptosporidium infection is not found in the normal popu-lation of Nanjing City,but the Cryptosporidium infection is found in the chronic diarrhea patients. The results imply that we should strengthen the detection of Cryptosporidium in the chronic diarrhea patients,so as to provide the evidence for improving the diagnosis and treatment of cryptosporidiosis.

Diagnosis of active tuberculosis disease: From microscopy to molecular techniques.

Methods used for the laboratory diagnosis of tuberculosis are continually evolving in order to achieve more rapid, less expensive, and accurate results. Acid-fast staining and culture for mycobacteria remain at the core of any diagnostic algorithm. Following growth in culture, molecular technologies such as nucleic acid hybridization probes, MALDI-TOF MS, and DNA sequencing may be used for definitive species identification. Nucleic acid amplification methods allow for the direct detection of Mycobacterium tuberculosis complex within respiratory specimens without relying on culture growth, leading to more rapid diagnoses and appropriate patient care.

A Comparative Study on Four Methods in the Diagnosis of Tuberculosis / 现代检验医学杂志

Objective To evaluate the value of four methods in diagnosis of pulmonary tuberculosis,including T-SPOT,fluo-rescent PCR,anti-TB antibody test,and acid-fast staining.Methods Retrospective analysis of 530 cases between January 2012 and December 2014 who had taken four methods,and calculated the sensitivity specificity,positive predictive value,neg-ative predictive value,Kappa value,Youden index,positive likelihood ratio,negative likelihood ratio,Pairedχ2 test.Consider clinical diagnosis as the gold standard.Results The sensitivity,negative predictive value,Youden index of T-SPOT were 95.90%,97.29%,0.82,respectively,and all of these were the highest.The negative likelihood ratio of T-SPOT was 0.05, which was the lowest.Misdiagnosis rate of PCR was 87.18%,which was the highest.Positive likelihood of anti-TB antibody test was the lowest,6.48,while other indicators were no advantage.The specificity,positive predictive value and positive likelihood ratio of acid-fast staining were 99.70%,98.90%,153.83,respectively,and the three of these were highest.Pair-wise comparison between the four methods were significantly different.Conclusion The T-SPOT and acid-fast staining can be used as important methods,and the anti-TB antibody can provide result quickly,and PCR method is more suitable for ex-amination of sterile body fluids.

Detection of Mycobacterium avium subspecies paratuberculosis infection in two different camel species by conventional and molecular techniques.

Paratuberculosis (John's disease) is infectious and chronically progressive granulomatous disease which affects domestic and wild ruminants. The causative agent is Mycobacterium avium paratuberculosis (MAP), a slow growing mycobactin dependent acid-fast bacillus. We investigated the detection and frequency of MAP in apparently healthy dromedary and Bactrian camels by insertion sequence 900 (IS900) polymerase chain reaction (PCR) and acid fast staining of fecal samples in Iran. Acid fast staining results showed that 6/50 (12.0%) samples of dromedary camels and 4/26 (15.3%) samples of Bactrian camels were suspected to MAP. Although the percentage of positivity for PCR assay of fecal dromedary camel was 8.0%, no bands corresponding to MAP detect in all samples of Bactrian camels. In conclusion, Although the incidence of MAP infection was low, further studies should be conducted to get more information on MAP infection in camel population, especially in areas where camels are close to other ruminants such as dairy cow, sheep and goat.

Evaluation on effect of fluorescent staining and Ziehl-Neelsen staining methods for detecting Mycobacterium tuberculosis / 国际检验医学杂志

Objective To compare the results difference between the fluorescence staining and the acid fast staining (Ziehi‐Neelsen staining) methods in the detection of Mycobacterium tuberculosis ,and to compare the effects of the methylene blue solu‐tion ,Haris hematoxylin solution and potassium permanganate liquid as the redyeing reagents of the fluorescence staining method . Methods 198 sputum specimens collected from the patients with suspected tuberculosis symptoms and were performed the Ziehi‐Neelsen staining and the fluorescence staining respectively For comparing the difference in the detecting rate of Mycobacterium tu‐berculosis between the two kinds of method .The fluorescence staining adopted 0 .3% methylene blue solution ,0 .5% Haris hema‐toxylin solution and 0 .5% potassium permanganate solution as the redyeing reagents for comparing the effects of the fluorescence microscopic examination among different redying reagents .Results The detection rate of Mycobacterium tuberculosis was 66 .67%(132/198) for the Ziehi‐Neelsen staining ,94 .9% (188/198) for the fluorescence stainings and 94 .95% (188/198) for the methyl‐ene blue staining ,in which the detection rate of methylene blue redyeing was 94 .95% ,which of hematoxylin redyeing was 94 .44%(187/198) and which of potassium permanganate redyeing was 94 .44 (187/198) ,the differences among them were statistically sig‐nificant(P< 0 .05) .Conclusion The fluorescent staining method has the higher positive detection rate of Mycobacterium tubercu‐losis than the Ziehl‐Neelsen staining method ,in which 0 .3% methylene blue solution is a good background quenching redyeing solu‐tion .

Clinical application and comparison of rapid and accurate identification of mycobacterium by gene chip microarray and smear acid-fast staining / 国际检验医学杂志

Objective To compare the clinical effect of application of gene chip microscopy technique for rapid identification of Mycobacterium and classic smear acid-fast staining,and to assess the advantages and disadvantages of the wo methods.Methods From 201 1 to 2014,gene chip microarray and smear acid fast staining were used to identify the mycobacterium tuberculosis in speci-mens suspicious of the infection from all the general hospitals of Shenzhen city.Chi-square test was used to compare the positive rates of the two methods.Results A total of 2 481 specimens were collected from clinic.With smear acid-fast staining technique, the positive specimens of 1 93 cases werefound and the positive rate was 7.8%.Meanwhile,31 7 positive samples were detected by the technology of gene chip microarray,and the positive rate was 12.8%.The positive rate of Gene chip microarray technology was higher than that of the smear acid fast staining,and there was significant difference between them (P < 0.05 ).The 31 7 positive samples identified by Gene chip microarray,included 263 cases of Mycobacterium tuberculosis,27 cases of Mycobacterium absces-sus,18 cases of Mycobacterium intracellulare,3 cases of Mycobacterium gastric uLcer,3 cases of Mycobacterium avium,1 case of Mycobacterium Gordonae,1 case of Mycobacterium marinum and 1 case of Mycobacterium Kansas.Conclusion The gene chip mi-croarray technology is fast,accurate,and its positive rate is higher than that of smear acid-fast staining technique.Classification and identification of Mycobacterium is very helpful for clinical individualized treatment of anti mycobacterium infection.