Actualización en la detección de la tuberculosis a través de pruebas moleculares / An update on the detection of tuberculosis through molecular testing

La tuberculosis es una enfermedad infecciosa respiratoria que afecta a un tercio de la población mundial y es una amenaza significativa para la salud global. La detección de la tuberculosis de manera temprana es crucial para un tratamiento eficaz y prevenir su propagación. Una solución para mejorar el diagnóstico y abordar la resistencia a los medicamentos antituberculosos es el uso de pruebas moleculares de alto rendimiento para la identificación del Mycobacterium tuberculosis y su susceptibilidad. Este estudio de revisión narrativa busca describir las generalidades, la eficacia, la sensibilidad, las ventajas y las limitaciones de las principales pruebas moleculares; Truenat® MTB, MTB plus y MTB-RIF, Abbott RealTime MTB y MTB RIF/INH en el sistema m2000sp y m2000rt y FluoroType MTBDR, además, de compararlas con GeneXpert MTB/RIF o Xpert Ultra, utilizadas para la detección del patógeno resistente a medicamentos tuberculosos. Estas pruebas utilizan diversas técnicas para la detección del ADN del Mycobacterium tuberculosis y la cuantificación de la carga bacteriana con alta sensibilidad y especificidad, resultados rápidos, reducción de los errores humanos, así como la detección temprana de cepas drogo-resistentes. A pesar de que requieren infraestructura especializada y competencias profesionales para su implementación, representan avances significativos con el potencial de mejorar la atención sanitaria y la gestión de la tuberculosis. Estas pruebas moleculares, comparadas con el GeneXpert, son una alternativa viable, aunque esta última tecnología sigue siendo la preferida en áreas con recursos limitados

Tuberculosis is a respiratory infectious disease that affects one third of the world's population and is a significant threat to global health. Detecting tuberculosis early is crucial for effective treatment and preventing its spread. One solution to improve diagnosis and address antituberculosis drug resistance is the use of high-throughput molecular tests for the identification of Mycobacterium tuberculosis and its susceptibility. This narrative review study seeks to describe the generalities, efficacy, sensitivity, advantages and limitations of the main molecular tests: Truenat® MTB, MTB plus and MTB-RIF, Abbott RealTime MTB and MTB RIF/INH on the m2000sp and m2000rt system and FluoroType MTBDR, and to compare them with GeneXpert MTB/RIF or Xpert Ultra, used for the detection of the tuberculosis drug-resistant pathogen. These tests use various techniques for the detection of Mycobacterium tuberculosis DNA and quantification of bacterial load with high sensitivity and specificity, rapid results, reduction of human error, as well as early detection of drug-resistant strains

Síndrome medular completo secundario a mielitis infecciosa causada por Mycobacterium tuberculosis. Informe de caso / Central spinal cord syndrome secondary to infectious myelitis caused by Mycobacterium Tuberculosis

Introducción: La tuberculosis, un problema de salud pública a nivel mundial que afecta a la población. Causada por Mycobacterium tuberculosis, se transmite a través de gotas de saliva y la presentación extrapulmonar afecta 5­45% de la población infectada. La tuberculosis de columna vertebral es una enfermedad crónica y progresiva, secundaria a la diseminación de un foco primario, que puede pasar desapercibido. Descripción del caso: Paciente masculino con fiebre de 4 días de evolución, nocturna, con diaforesis, debilidad de fuerza de miembros superiores e inferiores bilateral; miembros inferiores con (0) fuerza y miembros superiores (1), ausencia de reflejos osteotendinosos en ambas extremidades, nivel sensitivo C4, con ausencia de control de esfínteres. El diagnóstico se estableció mediante toracocentesis diagnóstica, realizando GeneXpert y cultivo de líquido pleural, positivo para tuberculosis y negativo para micobacteria resistente; resonancia magnética con gadolinio identifica área hiperintensa cervical medular de C2 hasta C4, produciendo aumento en el diámetro de la médula espinal. Se inició tratamiento tetraconjugado: isoniazida, rifampicina, pirazinamida y etambutol. Cuatro semanas posteriores al comienzo de tratamiento paciente presenta (2) fuerza en miembros inferiores y (3) en miembros superiores, continuando con terapia física. Conclusión: la tuberculosis se presenta de maneras muy diversas y se debe tener presente en pacientes jóvenes sin antecedentes personales patológicos que presenten cuadros de fiebre como uno de los primeros síntomas. Pruebas diagnósticas serológicas, GeneXpert y de imagen como la resonancia magnética ayudan en establecer un diagnóstico. La tuberculosis debe considerarse siempre en un proceso infeccioso que se desconoce etiología...(AU)

Tuberculosis extrapulmonar, la gran simuladora / Extrapulmonary tuberculosis, the great pretender

La tuberculosis es una enfermedad infecciosa considerada un problema de salud pública ya que constituye una de las principales causas de morbimortalidad a nivel mundial; su forma clínica más frecuente es la tuberculosis pulmonar, sin embargo, esta enfermedad también puede afectar estructuras extrapulmonares cuyo diagnóstico generalmente es tardío debido a que los síntomas y signos son inespecíficos. En este artículo se presenta un caso clínico de tuberculosis extrapulmonar (miliar, sistema nervioso central e intestinal) en el Hospital Universitario de Puebla (AU)

Tuberculosis is an infectious disease considered a public health problem since it is one of the main causes of morbidity and mortality worldwide; the most common clinical form is pulmonary tuberculosis; however, this disease can also affect extrapulmonary structures whose diagnosis is generally late because the symptoms and signs are nonspecific. This article presents a clinical case of extrapulmonary tuberculosis (miliary, central nervous system and intestinal) at the University Hospital of Puebla (AU)

Utilidad de GeneXpert MTB/RIF en el diagnóstico de tuberculosis frente a la baciloscopia / Utility of GeneXpert MTB/RIF in the Diagnosis of Tuberculosis Compared to Sputum Smear Microscopy

Introducción: La implementación de un método diagnóstico adecuado y eficiente es crucial para la detección temprana de la tuberculosis. Esto no solo permite un control efectivo de la enfermedad para evitar su transmisión y progresión hacia estadios más graves, además previene el desarrollo de resistencia a los fármacos en los pacientes.Objetivo: Evaluar la utilidad de la prueba molecular GeneXpert MTB/RIF en el diag-nóstico de Mycobacterium tuberculosis complex, en comparación con la bacilos-copia, utilizando el cultivo como referencia.Material y Métodos: Se realizó un estudio descriptivo, observacional y no expe-rimental de corte transversal, se incluyeron 253 muestras de pacientes de ambos sexos y de variados rangos de edad, que fueron evaluadas mediante baciloscopia, GeneXpert MTB/RIF y cultivo. El estudio se centró en muestras procesadas en un Hospital público de la ciudad de Quito durante el período de enero de 2021 a mayo de 2022Resultados: La prueba molecular GeneXpert MTB/RIF mostró una sensibilidad del 94,7% y una especificidad del 93,9% para el diagnóstico de Mycobacterium tu-berculosis complex. Además, se identificó un caso de resistencia a la rifampicina.Conclusión: Este estudio confirma la eficacia de la prueba molecular GeneXpert MTB/RIF sobre la baciloscopia para el diagnóstico oportuno de Mycobacterium tu-berculosis complex. Sin embargo, es esencial considerar las diversas condiciones de las muestras y pacientes para optimizar la precisión diagnóstica

Introduction: Implementing an appropriate and efficient diagnostic method is cru-cial for the early detection of tuberculosis. This not only allows for effective control of the disease to prevent its transmission and progression to more severe stages but also prevents the development of drug resistance in patients.Objective: To evaluate the utility of the GeneXpert MTB/RIF molecular test in diag-nosing Mycobacterium tuberculosis complex, compared to sputum smear micros-copy, using culture as the reference. Material and Methods: A descriptive, observational, and non-experimental cross-sectional study was conducted, including 253 samples from patients of both sexes and various age ranges, which were assessed using sputum smear micros-copy, GeneXpert MTB/RIF, and culture. The study focused on samples processed at a Quito ́s Public Hospital during the period from January 2021 to May 2022.Results: The GeneXpert MTB/RIF molecular test showed a sensitivity of 94.7% and a specificity of 93.9% for the diagnosis of Mycobacterium tuberculosis com-plex. Additionally, a case of resistance to rifampicin was identified.Conclusion: This study confirms the effectiveness of the GeneXpert MTB/RIF mo-lecular test over sputum smear microscopy for the timely diagnosis of tuberculosis. However, it is essential to consider the diverse conditions of the samples and pa-tients to optimize diagnostic accuracy

Tuberculosis abdominal en pediatría: presentación de un caso / Abdominal tuberculosis in pediatrics: A case report

La tuberculosis es una enfermedad infectocontagiosa cuya forma de presentación más frecuente es la pulmonar; la afectación abdominal es poco frecuente, por lo que su diagnóstico continúa siendo un desafío. Las manifestaciones clínicas de la tuberculosis abdominal así como sus hallazgos en el examen físico suelen ser inespecíficos y, en muchas ocasiones, similares a los de otras patologías, por lo que es fundamental considerarla entre los diagnósticos diferenciales. Se presenta el caso clínico de un paciente de sexo masculino, de 15 años de edad, hospitalizado por un síndrome febril prolongado asociado a dolor abdominal, diarrea, sudoración nocturna y pérdida de peso

Tuberculosis is an infectious disease which most commonly compromises the respiratory system, whereas abdominal involvement is rare, thus its diagnosis is a challenge. The clinical manifestations of abdominal tuberculosis as well as its physical examination findings are usually non-specific and, frequently, similar to those of other diseases, so it is critical to consider abdominal tuberculosis among the differential diagnoses. Here we report the clinical case of a 15-year-old male patient hospitalized for a prolonged febrile syndrome associated with abdominal pain, diarrhea, night sweats, and weight loss.

Mycobacterium tuberculosis resistente a Rifampicina en la provincia de El Oro-Ecuador / Rifampicin-resistant mycobacterium tuberculosis in the province of Oro-Ecuador / Mycobacterium tuberculosis resistente à Rifampicina na província de Oro-Equador

Introducción. La tuberculosis es una de las enfermedades infecciosas más antiguas y comunes, causada por una bacteria en forma de bastón, o bacilo, llamada Mycobacterium tuberculosis. Esta enfermedad es tratable con antibióticos, los cuales se prescriben durante meses debido a la lenta tasa de crecimiento de las bacterias. Entre los fármacos utilizados la Rifampicina se utiliza terapéuticamente para combatir esta enfermedad, sin embargo, algunos pacientes pueden presentar o desarrollar resistencia a este antibiótico, por lo que, es importante completar el tratamiento para evitar el desarrollo de bacterias farmacorresistentes y la re ocurrencia de la enfermedad. Objetivo. Caracterizar el Mycobacterium tuberculosis resistente a Rifampicina en la provincia de El Oro. Materiales y métodos. Se realizó un estudio con un enfoque cuantitativo, de tipo descriptivo y no experimental. La muestra se obtuvo de la base de datos del laboratorio clínico del área de micobacterias del Hospital Teófilo Dávila en el período 2019 - 2022, quienes luego de aplicar la prueba molecular rápida GeneXpert MTB/RIF o ULTRA determinaron que 48 pacientes presentaron resistencia a la Rifampicina en el tratamiento de la Mycobacterium tuberculosis. Resultados. El reporte del laboratorio evidenció que en el año 2022 se estableció el mayor número de casos de resistencia a Rifampicina para el tratamiento de Mycobacterium tuberculosis, alcanzando el 33,3%; el grupo etario de mayor afectación fue el adulto joven (20 a 49 años) con un 52,1%, y con una frecuencia elevada de 66,7% el sexo masculino. La comorbilidad con mayor predominio estuvo en los pacientes diagnosticados Diabetes Mellitus tipo 2 con un 27,1% y la condición de ingreso de pacientes con resistencia a Rifampicina, son de nuevos casos con 75%. Conclusiones. En la provincia de El Oro, entre el año 2019 - 2022 se presentaron 48 casos resistentes al antibiótico Rifampicina en el tratamiento de la TB, entre ellos el 75% corresponden a una resistencia inicial, es decir, pacientes que no fueron tratados contra la enfermedad, el otro 25% engloba a aquellos pacientes que recayeron en la enfermedad, fracaso o perdida de seguimiento por parte del laboratorio de vigilancia, área de micobacterias del Hospital Teófilo Dávila.

Introduction. Tuberculosis is one of the oldest and most common infectious diseases, caused by a rod-shaped bacterium, or bacillus, called Mycobacterium tuberculosis. This disease is treatable with antibiotics, which are prescribed for months due to the slow growth rate of the bacteria. Among the drugs used, Rifampicin is used therapeutically to combat this disease, however, some patients may present or develop resistance to this antibiotic, therefore, it is important to complete the treatment to avoid the development of drug-resistant bacteria and the reoccurrence of the disease. Objective. To characterize the Mycobacterium tuberculosis resistant to Rifampicin in the province of El Oro. Materials and methods. A quantitative, descriptive and non-experimental study was carried out. The sample was obtained from the database of the clinical laboratory of the mycobacteria area of the Teófilo Dávila Hospital in the period 2019 - 2022, who after applying the rapid molecular test GeneXpert MTB/RIF or ULTRA determined that 48 patients presented resistance to Rifampicin in the treatment of Mycobacterium tuberculosis. Results. The laboratory report showed that in the year 2022 the highest number of cases of resistance to Rifampicin for the treatment of Mycobacterium tuberculosis was established, reaching 33.3%; the age group most affected was young adults (20 to 49 years) with 52.1%, and with a high frequency of 66.7% in the male sex. The most prevalent comorbidity was in patients diagnosed with Diabetes Mellitus type 2 with 27.1% and the condition of admission of patients with resistance to Rifampicin, are new cases with 75%. Conclusions. In the province of El Oro, between 2019 - 2022 there were 48 cases resistant to the antibiotic Rifampicin in the treatment of TB, among them 75% correspond to an initial resistance, that is, patients who were not treated against the disease, the other 25% encompasses those patients who relapsed in the disease, failure or loss of follow-up by the surveillance laboratory, mycobacteria area of the Teófilo Dávila Hospital.

Introdução. A tuberculose é uma das doenças infecciosas mais antigas e mais comuns, causada por uma bactéria em forma de bastonete, ou bacilo, chamada Mycobacterium tuberculosis. A doença pode ser tratada com antibióticos, que são prescritos por meses devido à lenta taxa de crescimento da bactéria. Entre os medicamentos utilizados, a rifampicina é usada terapeuticamente para combater essa doença; no entanto, alguns pacientes podem desenvolver resistência a esse antibiótico, por isso é importante concluir o tratamento para evitar o desenvolvimento de bactérias resistentes aos medicamentos e a recorrência da doença. Objetivo. Caracterizar o Mycobacterium tuberculosis resistente à rifampicina na província de El Oro. Materiais e métodos. Foi realizado um estudo quantitativo, descritivo e não experimental. A amostra foi obtida do banco de dados do laboratório clínico da área de micobactérias do Hospital Teófilo Dávila no período de 2019 a 2022, que, após aplicar o teste molecular rápido GeneXpert MTB/RIF ou ULTRA, determinou que 48 pacientes apresentavam resistência à rifampicina no tratamento de Mycobacterium tuberculosis. Resultados. O laudo laboratorial demonstrou que o maior número de casos de resistência à Rifampicina para o tratamento do Mycobacterium tuberculosis se estabeleceu em 2022, atingindo 33,3%; a faixa etária mais afetada foi a de adultos jovens (20-49 anos) com 52,1%, e com alta frequência de 66,7% no sexo masculino. A comorbidade mais prevalente foi em pacientes diagnosticados com Diabetes Mellitus tipo 2 com 27,1% e a condição de admissão de pacientes com resistência à Rifampicina, são casos novos com 75%. Conclusões. Na província de El Oro, entre 2019 e 2022, houve 48 casos resistentes ao antibiótico Rifampicina no tratamento da TB, entre eles 75% correspondem a uma resistência inicial, ou seja, pacientes que não foram tratados contra a doença, os outros 25% incluem aqueles pacientes que recaíram na doença, falha ou perda de monitoramento pelo laboratório de vigilância, área de micobactérias do Hospital Teófilo Dávila.

Mycobacterium tuberculosis y virus linfotrópico humano, una co-infección fatal / Mycobacterium tuberculosis and human lymphotropic virus, a fatal co-infection

La tuberculosis es una infección de alta incidencia en Latinoamérica. Su presentación como infección activa está determinada por factores de riesgo del hospedero. Comunicamos el caso clínico de una mujer joven que presentó una forma grave de tuberculosis pulmonar. Al explorar sus factores de riesgo se confirmó un estado de inmunosupresión profundo, causado por un linfoma de células T, asociada a una co-infección por virus linfotrópico T humano tipo 1. Se destacan los aspectos microbiológicos y de pronóstico de la co-infección de Mycobacterium tuberculosis y HTLV-1

Tuberculosis is a high-incidence infection in Latin America. Its presentation as an active infection is determined by risk factors in the host. We report the case of a young woman who presented a severe form of pulmonary tuberculosis. When exploring her risk factors, a profound state of immunosuppression was found, caused by T-cell lymphoma, associated with co-infection with human lymphotropic virus. Microbiological and prognostic aspects of Mycobacterium tuberculosis and HTLV-1 co-infection are highlighted.

Ag85B with c-di-AMP as mucosal adjuvant showed immunotherapeutic effects on persistent Mycobacterium tuberculosis infection in mice

Tuberculosis (TB), caused by Mycobacterium tuberculosis, remains the leading cause of mortality by a single infectious agent in the world. M. tuberculosis infection could also result in clinical chronic infection, known as latent TB infection (LTBI). Compared to the current limited treatment, several subunit vaccines showed immunotherapeutic effects and were included in clinical trials. In this study, a subunit vaccine of Ag85B with a novel mucosal adjuvant c-di-AMP (Ag85B:c-di-AMP) was delivered intranasally to a persistent M. tuberculosis H37Ra infection mouse model, which also presented the asymptomatic characteristics of LTBI. Compared with Ag85B immunization, Ag85B:c-di-AMP vaccination induced stronger humoral immune responses, significantly higher CD4+ T cells recruitment, enhanced Th1/Th2/Th17 profile response in the lung, decreased pathological lesions of the lung, and reduced M. tuberculosis load in mice. Taken together, Ag85B:c-di-AMP mucosal route immunization provided an immunotherapeutic effect on persistent M. tuberculosis H37Ra infection, and c-di-AMP, as a promising potential mucosal adjuvant, could be further used in therapeutic or prophylactic vaccine strategies for persistent M. tuberculosis infection as well as LTBI.

Masa nasal como único compromiso extrapulmonar de infección por M. Tuberculosis / Nasal mass as the only extrapulmonary compromise of M. Tuberculosis infection / Massa nasal como único compromisso extrapulmonar da infecção por M. Tuberculose

La tuberculosis aún es un problema de salud pública mundial. La infección causada por Mycobacterium tuberculosis se manifiesta de forma principal a nivel pulmonar. Sin embargo, alrededor del 20 % de los casos se presentan en otras localizaciones anatómicas y solo el 2 % tiene afectación del tracto respiratorio superior. Se presenta el caso de una mujer de 75 años, reconsultante al servicio de otorrinolaringología por epistaxis, lesiones postillosas en cavidad nasal y hallazgo de masa nasal. Posterior a la resección quirúrgica de la lesión, se logró la comprobación microbiológica de infección por M. tuberculosis. Se realizan estudios para descartar compromiso pulmonar y de otras localizaciones. Posterior al inicio de tratamiento antituberculoso se logró resolución completa de la lesión y no recurrencia de los síntomas. Las formas extrapulmonares de la infección por M. tuberculosis y, en especial las que afectan la región de la cabeza y el cuello, requieren un alto índice de sospecha para su diagnóstico. Los métodos de diagnóstico como la prueba de PCR y los cultivos de tejidos permiten un óptimo inicio del manejo médico de acuerdo con la epidemiología local y las condiciones del paciente.

Tuberculosis is still a global public health burden. Infection caused by the bacillus Mycobacterium tuberculosis (M. Tuberculosis) manifests mainly in the lungs. However, around 20 % of cases occur in other anatomical locations and only 2 % have upper respiratory tract involvement. We present the case of a 75-year-old female patient, who returned to the otorhinolaryngology service due to epistaxis and postillomous lesions in the nasal cavity with a finding of a nasal mass. After surgical resection of the lesion, microbiological confirmation of M. tuberculosis infection is achieved. Studies are performed to rule-out lung involvement, as well as other locations. After the initiation of tuberculosis treatment, complete resolution of the lesion and no recurrence of symptoms is documented. Extrapulmonary forms of M. tuberculosis infection, and especially those involving the head and neck region, require a high index of suspicion for their diagnosis. Diagnostic methods such as PCR testing and tissue cultures allow optimal initiation of medical management according to local epidemiology and patient conditions.

A tuberculose ainda é um problema de saúde pública global. A infecção causada pelo Mycobacterium tuberculoses manifesta-se principalmente nos pulmões. Entretanto, cerca de 20% dos casos ocorrem em outras localizações anatômicas e apenas 2% apresentam comprometimento do trato respiratório superior. É apresentado o caso de uma mulher de 75 anos que retornou ao serviço de otorrinolaringologia por quadro de epistaxe, lesões com crostas em cavidade nasal e descoberta de massa nasal. Após ressecção cirúrgica da lesão, foi realizada verificação microbiológica de infecção por M. tuberculoses. Estudos são realizados para descartar envolvimento pulmonar e otras localizações. Após início do tratamento antituberculoso, houve resolução completa dalesão e não houve recidiva dos sintomas. As formas extrapulmonares da infecção por M. tuberculoses, especialmente aquelas que acometem a região de cabeça e pescoço, requerem alto índice de suspeita para diagnóstico. Métodos de diagnóstico, como testes de PCR e culturas de tecidos, permitem o início ideal do tratamento médico de acordó com a epidemiologia local e as condições do paciente.

Prokaryotic expression and functional analysis of molecular chaperone Acr2 protein of Mycobacterium tuberculosis / 中国生物制品学杂志

@#Objective To express the molecular chaperone Acr2 protein of Mycobacterium tuberculosis(Mtb)in E.coli and analyze the function. Methods The recombinant plasmid pET-28a-Acr2 was transformed into competent E. coli BL21(DE3),and induced by IPTG. The expressed His-Acr2 protein was purified by Ni-NTA chromatography and SuperdexTM200 10/300 GL gel filtration chromatography to obtain Acr2 protein. The Acr2 protein was refolded by spontaneous refolding and reassembly after thermal denaturation(100 ℃ for 15 min)and chemical denaturation(8 mol/L urea,37 ℃ for 4 h).The secondary structure of Acr2 protein before and after denaturation-renaturation was detected by circular dichroism spectroscopy and non-denaturing SDS-PAGE,and the molecular chaperone function of Acr2 protein in vitro was detected by substrate binding assay. Results The purified Acr2 protein had the relative molecular mass of about 232 000,the purity of over 90%,and the concentration of about 2 mg/mL,which recovered its natural secondary structure after denaturationrenaturation,and formed stable complexes with the denatured malate dehydrogenase(MDH)at 48 ℃. Conclusion The Acr2protein can restore its natural molecular conformation with molecular chaperone activity in vitro after denaturation-renaturation treatment,providing a new strategy for the preparation of Mtb protein antigen with natural activity.

Development and verification of a reversed phase high-performance liquid chromatography method for purity determination of recombinant Mycobacterium tuberculosis Ag85b protein stock solution / 中国生物制品学杂志

@#Objective To develop and verify a reversed phase high-performance liquid chromatography method for the determination of the purity of recombinant Mycobacterium tuberculosis(Mtb)Ag85b protein stock solution.Methods Fourfactor,three-level orthogonal test was designed,with the area,trailing factor,peak area and peak area RSD as the evaluation indexes to explore the optimal detection conditions. The methodology verification of specificity,linear range,precision and durability was conducted in accordance with the general principles of Chinese Pharmacopoeia(Volume Ⅳ,2020 edition)9101.Results The results of all the evaluation indexes were good when the elution ratio of organic phase was30% ～ 95%,the detection temperature was 35 ℃,the sample volume was 3 μg,and the elution time of 95% organic phase was 15 min. The method had the linear correlation coefficient(R2)of 0. 998 5,the linear range of 1. 8 ～ 4. 2 μg,the reproducibility RSD of 0. 01%,and the intermediate precision RSD of 0. 16%,with good durability under slight changes of column temperature and flow rate.Conclusion The reversed phase high-performance liquid chromatography method for the purity determination of recombinant Mtb Ag85b protein stock solution was developed,which has good specificity,precision and durability,and can be used for the quality control of recombinant Mtb Ag85b protein stock solution.

Guideline for clinical diagnosis and treatment of tuberculosis in kidney transplant recipients in China (2023 edition) / 器官移植

This guideline aims to provide comprehensive and practical guidance for clinical management of tuberculosis in kidney transplant recipients. First, it summarizes the particularity of tuberculosis in kidney transplant recipients, and highlights the high incidence and diverse clinical manifestations. To better understand the patients' conditions, relevant assessment of tuberculosis is recommended before kidney transplantation. Extensive attention should be paid to the monitoring of tuberculosis after kidney transplantation. Regarding the diagnosis, the guideline explicitly introduces common diagnostic approaches for tuberculosis, and evaluates the applicability in kidney transplant recipients. After the diagnosis is confirmed, it discusses how to balance the treatment and rejection of tuberculosis under the background of immunosuppressants, and focuses upon the potential drug interaction. In terms of prevention, it emphasizes the screening of tuberculosis prior to kidney transplantation. This guideline is designed to deepen the understanding of medical staff for tuberculosis management in kidney transplant recipients, promote more effective clinical practice and improve the quality of life of the recipients.

Comparative study of drug susceptibility testing and whole genome test testing anti-tuberculosis drug resistance / 国际检验医学杂志

Objective To compare the categorical agreement between drug susceptibility testing(DST)and whole genome sequencing(WGS)for the detection of drug resistance in Mycobacterium tuberculosis(MTB),and to explore the characteristics of WGS for MTB drug resistance detection.Methods A total of 71 MTB clinical isolates retained in West China Hospital of Sichuan University from 2018 to 2020 were included in this study.The MTB strains were tested for resistance to 14 anti-tuberculosis drugs,including Isoniazid(INH),Rifampicin(RIF),Rifabutin(RFB),Ethambutol(EMB),Streptomycin(SM),Moxifloxacin(MFX),Ofloxacin(OFX),Levofloxacin(LFX),Amikacin(AMK),Kanamycin(KAN),Capreomycin(CPM),Para-aminosalicylic acid(PAS),Ethionamide(ETH)and Clofazimine(CLO),using both DST(colorimetric redox indicator meth-od)and WGS methods.Kappa test was performed to analyze the results of drug resistance detection for both methods.Results Based on DST and WGS methods to detect anti-tuberculosis drug resistance in seventy-one MTB clinical isolates,the results showed that the agreement rate of RIF,RFB,SM,MFX,OFX and LFX ex-ceeded 90.00％,and the kappa values were all greater than 0.80,with near perfect agreement;The agreement rates of INH and EMB were 84.51％and 81.69％,and Kappa values were 0.68 and 0.54,respectively,with fair agreement.No more than two drug resistant MTB strains of AMK and KAN were detected by both meth-ods,and the resistance rate was less than 3.00％.The agreement rates of CPM,ETH,PAS,and CLO ranged from 61.97％to 91.55％,and the Kappa values were less than 0.40,with slight or fair agreement.Conclusion There are differences in the ability of WGS to detect resistance to various anti-tuberculosis drugs,and it is more effective in detecting resistance to six anti-tuberculosis drugs,including RIF,RFB,SM,MFX,OFX and LFX,while there are still certain differences in detecting resistance to other anti-tuberculosis drugs compared with DST.It is necessary to further clarify the detailed resistance mechanisms of relevant anti-tu-berculosis drugs and to explore the standardization of WGS for drug resistance detection.

Research Progress of Non-coding RNA in Autophagy of Macrophages During Mycobacterium tuberculosis Infection / 华中科技大学学报(医学版)

Tuberculosis(TB)is an infectious disease caused by Mycobacterium tuberculosis(MTB).Autophagy plays an im-portant role in eliminating MTB which can interfere with host autophagy through a variety of mechanisms,therefore,escape the killing of macrophages to survive and reproduce in the host cells for a long time.There are many kinds of differential expressed non-coding RNAs(ncRNAs)in macrophages upon MTB infection.These ncRNAs can affect the elimination of MTB by regula-ting the expression of autophagy-related genes in multiple processes of autophagy.Elucidating these regulatory networks are of great significance for the development of new anti-TB drugs.Therefore,the known mechanisms of ncRNAs that regulates the autophagy of MTB infected macrophages were summarized,and the process of autophagy and the main regulatory functions of ncRNAs were introduced in this review,in order to provide new perspectives for the pathogenesis of TB and the development of anti-TB drugs.

Construction of eukaryotic expression vector for bacteriophage D29 LysinB/Holin and analysis of its bactericidal activity / 中华微生物学和免疫学杂志

Objective:To construct a eukaryotic expression vector for bacteriophage D29 LysinB/Holin fusion protein and study its bactericidal efficacy against Mycobacterium tuberculosis ( Mtb) in a cell infection model. Methods:A recombinant plasmid pET32a-LysinB was constructed and induced to express LysinB. The polyclonal antibody against LysinB was prepared after the purification of LysinB. A recombinant plasmid pcDNA3.1(+ )-LysinB/Holin was constructed and transfected into mononuclear macrophages RAW264.7. After the expression of the prepared polyclonal antibody was identified, a cell infection model was established and the bactericidal efficacy of LysinB/Holin fusion protein was measured by acid-fast staining and colony counting.Results:The polyclonal antibody against LysinB was successfully prepared. The recombinant plasmid pcDNA3.1(+ )-LysinB/Holin could effectively express LysinB/Holin fusion protein in eukaryotic cells without inducing significant cytotoxicity. LysinB/Holin fusion protein was effective in killing Mtb in cells. Conclusions:The recombinant plasmid pcDNA3.1(+ )-LysinB/Holin has a better killing effect on intracellular Mtb without inducing obvious cytotoxicity against eukaryotic cells, showing a potential in the treatment of tuberculosis.

Mycobacterium tuberculosis extracellular vesicles: diagnostic, preventive and therapeutic potential / 中华检验医学杂志

By summarizing the biological formation and component function of Mycobacterium tuberculosis extracellular vesicles, this manuscript discusses the potential usage of Mycobacterium tuberculosis extracellular vesicles in diagnosis, prevention, and treatment of tuberculosis. It is concluded that both Mycobacterium tuberculosis membrane vesicles and exosomes secreted by the host contain the components of Mycobacterium tuberculosis to participate in immune regulation, and play different roles in the diagnosis, prevention, and treatment of tuberculosis. Mycobacterium tuberculosis extracellular vesicles contain abundant biomarkers and have natural immunogenicity, which are expected to become new targets for diagnosis, prevention, and treatment of tuberculosis.

The potential of a self-nano emulsifying drug delivery system for ethanol extract of red betel vine as an antibacterial agent against Mycobacterium tuberculosis and Pseudomonas aeruginosa

Aims@#Mycobacterium tuberculosis and Pseudomonas aeruginosa are still a problem in the world. This study was aimed to develop the self-nano emulsifying drug delivery system (SNEDDS) formulation of red betel vine leaf ethanol extract, which has the potential as an antibacterial agent against M. tuberculosis and P. aeruginosa. @*Methodology and results@#Red betel vines were extracted using the maceration method with 70% ethanol. SNEDDS ethanol extract was prepared using high-energy and low-energy methods, while the formulation was evaluated in the form of % transmittance, particle size, polydispersion index and zeta potential to accelerated stability tests. The SNEDDS formula of the extract, passing the stability test, was further tested for accelerated antibacterial activity. The presence of M. tuberculosis was tested using the resazurin microtiter assay (REMA) method, while P. aeruginosa was tested using the dilution method by determining the minimum bactericidal concentration (MBC) value. In this study, the SNEDDS formula of the extract, which included nanoemulsion, had a particle size of 16-17 nm (<200 nm), PI 0.52-0.55 D (<0.7 D), zeta potential -18.03-24.96 mV (<30 mV) and % transmittance of 94-96% (>90%). MBC of SNEDDS formula against M. tuberculosis becomes apparent at concentrations <1%, whereas in the case of P. aeruginosa, this phenomenon is observed at a concentration as low as 0.0312%. @*Conclusion, significance and impact of study: @#Research findings suggest that the SNEDDS formula of red betel vine leaf ethanol extract was stable and has the potential as an antibacterial agent against M. tuberculosis and P. aeruginosa.

Expert consensus on the bi-directional screening for Mycobacterium tuberculosis and human immunodeficiency virus / 上海预防医学

Tuberculosis （TB） and human immunodeficiency virus infection / acquired immune deficiency syndrome （HIV/AIDS） are both serious global public health threats. Early detection of infected persons and/or patients through TB/HIV bi-directional screening is crucial for prevention and control strategy in China and globally. In recent years， with the promotion and application of new TB and HIV detection technologies worldwide， TB/HIV bi-directional screening technologies and strategies have made remarkable changes. This expert consensus introduces the significance and challenges of TB/HIV bi-directional screening， summarizes important progress of research and applications， and makes recommendations on screening measures and procedures to further strengthen TB/HIV bi-directional screening in China.

Latest advances in the development of tuberculosis vaccines in global clinical trials: a review / 中国血吸虫病防治杂志

Tuberculosis (TB) remains one of the biggest infectious killers worldwide. Vaccine is the most satisfactory tool for prevention of TB; however, Bacillus Calmette-Guérin (BCG), the widely used vaccine in clinical for the prevention of TB, has limitations in protective effects. Development of novel TB vaccines is therefore of urgent need. Currently, there are 15 novel TB vaccine candidates in clinical trials, including live-attenuated vaccines, inactivated vaccines, subunit vaccines and viral-vectored vaccines, which open the door for the ultimate target of the End TB Strategy. This review summarizes the latest advances in the development of TB vaccines in global clinical trials, so as to provide insights into TB control.

Status quo and influencing factors of drug resistance of tuberculosis in Inner Mongolia / 安徽医科大学学报

Objective @#To investigate the status quo of drug resistance and influencing factors of tuberculosis in Inner Mongolia , and to provide reference for accurate prevention and control of drug - resistant tuberculosis .@*Methods @#Random sampling was used in this study . TB patients from Tuberculosis designated hospital in Inner Mongolia were included , according to the rules and drug - resistant strains were identified and tested according to relevant norms . Composition ratio or rate was calculated for statistical description , and Logistic regression model was used to analyze the influencing factors of drug resistance in TB patients .@*Results @#Among 1321 patients, there were 936 males and 385 females, with an average age of (52.65 ±18 .09) years . The rates of mono - resistant , multidrug - resistant (MDR) , extensively drug - resistant (XDR) and total drug resistance were 19.00% , 11.58% , 11.66% and 42.24% , respectively . The highest resistance rates were ob served for streptomycin ( 7.27% ) , isoniazid (4.69% ) , and isoniazid + streptomycin (4.47% ) . The drug resistance spectrum presented diversity and com plexity . Compared to females , males had a higher proportion of drug resistance , and the difference was statistically significant (P < 0.001) . The proportion of patients who were sensitive to anti - tuberculosis drugs increased with age (P < 0.05) . Among different age groups , the proportion of drug - resistant patients was higher in the 20 - 40 age group , 40 - 60 age group , and 60 and above age group compared to the 0 - 20 age group (P < 0.05) . Addi tionally , the proportion of drug - resistant patients was higher in the 20 - 40 age group and 40 - 60 age group com pared to the 60 and above age group (P < 0.05) . Moreover , the proportion of drug - resistant and multi - drug resistant patients was higher among patients undergoing retreatment compared to those undergoing initial treatment (P < 0.001) . Multivariate Logistic regression analysis showed that male gender (OR = 1.48 , 95% CI: 1.02 - 2.14) , age 20 - 40 years (OR = 2.64 , 95% CI: 1.05 - 6.60) , retreatment (OR = 2.34 , 95% CI: 1.70 - 3.22) , and outpatient follow-up (OR = 1.56 , 95% CI: 1.05 - 2.33) were independent risk factors for drug - resistant tuber culosis .@*Conclusion @#Inner Mongolia has a high prevalence of MDR and overall drug - resistant tuberculosis among patients . The drug resistance profile exhibits diversity and complexity . Risk factors that contribute to drug resistance include being male , aged between 20 and 40 , undergoing retreatment , and receiving outpatient follow - up . Therefore , it is necessary to further improve clinical diagnosis and treatment , promote rational use of first - line anti - tuberculosis drugs , prioritize individualized treatment , enhance health education , improve the medical insurance system , and optimize patient management approaches in order to enhance patient compliance .