Objective: To understand the recent transmission of Mycobacterium tuberculosis (MTB), and to identify the influencing factors of recent transmission among pulmonary tuberculosis (TB) patients in Jing'an district, Shanghai. Methods: The genotypes and drug resistances of MTB isolated from TB patients registered in the TB designated hospitals in Jing'an district during 2010-2015 were analyzed through 12-loci Mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR)(QUB11b, QUB18, Mtub21, Miru26, QUB26, Mtub04, Miru31, Miru40, VNTR2372, VNTR3820, 3232, 4120), and tested for drug susceptibility as well. With the results of field epidemiological investigation, univariate and multivariate analyses were performed to analyze the distribution of the clusters and influencing factors on recent transmission. Results: This study enrolled 80 TB patients, 23 (28.75%) had a resistance to at least one anti-TB drug, and the prevalence of multidrug-resistant tuberculosis (MDR-TB) was 16.25%. A total of 65 genotypes were identified with 58 (72.50%, 58/80) being unique and 7 clusters with 2-10 isolated in each cluster. The proportion of clustering was 27.50% (22/80). Results from the multivariate analysis revealed that multidrug- resistance (OR=35.799, 95%CI: 4.239-302.346) and having comorbidity with TB (OR=7.695, 95%CI: 1.421-41.658) were independently associated with the clustering, which suggesting a recent transmission. The field investigation to the clustered cases proved that the patients in two clusters had epidemiological links, one was between family members, and the other contained 10 MDR-TB patients with 9 knowing each other which have a definite connection and 1 having the possible connection with them. Conclusion: Recent transmission of tuberculosis happened among TB patients in Jing'an district, with high risks among the MDR-TB patients.
China ; Cluster Analysis ; Genotype ; Humans ; Mycobacterium tuberculosis/isolation & purification* ; Tuberculosis ; Tuberculosis, Multidrug-Resistant/transmission* ; Tuberculosis, Pulmonary/transmission*

China ; Humans ; Mycobacterium tuberculosis ; isolation & purification ; Quality Control ; Sputum ; microbiology ; Tuberculosis, Pulmonary ; microbiology

Nine proteins encoded by Mycobacterium tuberculosis RD1 region are important protective antigens that become absent in long passaging of Mycobacterium tuberculosis. They only exist in pathogenic Mycobacteria and are absent in Bacille Calmette-Guerin and environmental Mycobacteria. With good immunogenicities, they may play an important role in the diagnosis and prevention of Mycobacterium tuberculosis. This article reviews recent studies on using RD1-encoded proteins as antigens in the diagnosis of active tuberculosis and tuberculous pleurisy.
Antigens, Bacterial ; isolation & purification ; metabolism ; Humans ; Mycobacterium tuberculosis ; physiology ; Tuberculosis ; diagnosis

Diagnostic Tests, Routine ; methods ; Humans ; Mycobacterium tuberculosis ; isolation & purification ; Sensitivity and Specificity ; Tuberculosis, Pulmonary ; diagnosis ; microbiology

A femail patient was presented to our department for a dark red painless mass in the left auricle, which has progressively enlarged for one year. Pathological examination revealed granuloma change. Mycobacterium tuberculosis DNA test was positive. Tuberculin test revealed strong positive reaction. This case was diagnosed as tuberculous granuloma.
DNA, Bacterial ; isolation & purification ; Ear Auricle ; microbiology ; pathology ; Female ; Granuloma ; microbiology ; pathology ; Humans ; Mycobacterium tuberculosis ; isolation & purification ; Tuberculosis ; pathology

PURPOSE: Polymerase chain reaction (PCR) assay, introduced as a fast and sensitive diagnostic method, is useful in detecting Mycobacterium tuberculosis. The purpose of this study was to evaluate the usefulness of in-house PCR assay in the detection of Mycobacterium tuberculosis by comparing PCR results with conventional diagnostic techniques and Cobas Amplicor M. tuberculosis(TM) kit. MATERIALS and METHODS: We retrospectively assessed the diagnostic yield of in-house PCR method employed for the amplification IS6110 sequences in 2,973 specimens. We also compared in-house PCR with Cobas Amplicor M. tuberculosis(TM) kit in 120 specimens collected from June to July 2006. Routine acid-fast stain (AFS) and culture assay were also performed and analyzed. RESULTS: Of 2,973 cases, 2,832 cases (95.3%) showed consistent results between in house PCR, AFS and culture methods, whereas 141 (4.7%) displayed inconsistent results. The sensitivities, specificities, and positive and negative predictive values of each method were as follows: 77.5%, 99.7%, 95.5%, and 98.0%, respectively for PCR; 49.2%, 100%, 100%, and 95.7%, respectively, for AFS method; and 80.7%, 100%, 100%, and 98.3%, respectively, for culture assay. Consistent results between PCR and Cobas Amplicor M. tuberculosis(TM) kit were shown in 109 cases (90.8%). The sensitivities, specificities, and positive and negative predictive values of each method were as follows: 81.3%, 98.9%, 96.3%, and 93.5% respectively for PCR and 71.9%, 100%, 100%, and 90.7%, respectively, for Cobas Amplicor(TM) kit. CONCLUSION: In-house PCR and Cobas Amplicor(TM) kit show high sensitivity and specificity, and are reliable tests in the diagnosis of tuberculosis.
Humans ; Mycobacterium tuberculosis/*genetics/*isolation & purification ; Polymerase Chain Reaction/*methods ; Sensitivity and Specificity

Mycobacterium lentiflavum has recently been described as an emerging human pathogen without regard to the immune status of the host. We herein report on M. lentiflavum isolated from a respiratory specimen of a patient. Although the organism described in this case seems to be a colonizer of a lung destroyed by tuberculosis, the current methods for species identification of nontuberculous mycobacteria have to be re-evaluated so as not to underestimate these organisms.
Aged ; Humans ; Male ; Mycobacterium/genetics/*isolation & purification ; Tomography, X-Ray Computed ; Tuberculosis, Pulmonary/diagnosis/*microbiology/radiography

OBJECTIVEThis research was to establish a method for fast identification of mycobacteria in microtiter liquid culture and to evaluate its clinical value.

METHODS2-thiophenecarboxylic acid hydrazide (TCH) and paranitrobenzoic acid (PNB) at different concentrations were added into liquid culture in 96-well plate. Different mycobacterium standard strains were incubated in liquid culture with PNB and TCH for 7 to 10 days. According to the growth assay for 15 mycobacterium strains and 30 mycobacterium tuberculosis strains, the best PNB and TCH concentration were determined. A total of 424 clinical mycobacterium isolates were identified by microtiter liquid culture at the best PNB and TCH concentration. The results of microtiter liquid culture were compared with those of PCR and DNA sequencing.

RESULTSThe best concentration of PNB was 200 µg/ml in microtiter liquid culture. Compared with the results of PCR, the sensitivity and specificity for identification of mycobacterium tuberculosis complex in microtiter liquid culture were 97.8% (306/313) and 100.0% (107/107) respectively and those for non-tuberculosis mycobacteria in microtiter liquid culture were 100.0% (107/107) and 96.5% (306/317) respectively. The best concentration of TCH was 0.5 µg/ml. Compared with the results of PCR, the sensitivity of mycobacterium tuberculosis in microtiter liquid culture was 100.0% (305/305). The specificity remained under and more studies were needed.

CONCLUSIONIn microtiter liquid culture with PNB and TCH, mycobacteria can be identified in 7 to 10 days. The results were accurate and the process was simple without expensive equipments. This method meets clinical needs and can be used in all level hospitals in China.

OBJECTIVETo prepare reference samples of Mycobacterium tuberculosis culture filtrate protein-10 (CFP-10) and CFP10-streptavidin fusion proteins (CFP10/SA) for time-resolved fluoroimmunoassay (TRFIA).

METHODSThe CFP10 gene was amplified by PCR from Mycobacterium tuberculosis strain H37Rv and cloned into pET24b, pET24b-streptavidin (SA) or pET21a-SA expression vectors. The recombinant proteins CFP10, CFP10-SA and SA-CFP10 were expressed in Rosetta cells, purified via nickel affinity chromatography and refolded by dialysis. The sensitivity and stability of the resultant proteins as reference samples were evaluated by double-antibody sandwich TRFIA.

RESULTSCFP10-SA and SA-CFP10 fusion proteins were expressed as inclusion bodies, whereas CFP10 was expressed in a soluble form. The resultant purity of the 3 recombinant proteins all exceeded 95%. TRFIA results showed that CFP-SA fusion protein possessed the best sensitivity (0.02 µg/L) and stability.

CONCLUSIONThe reference samples of CFP10 for TRFIA detection have been successfully prepared and can be used in the development of a diagnostic kit for Mycobacterium tuberculosis.

Bacterial Typing Techniques ; China ; Humans ; Interspersed Repetitive Sequences ; Mycobacterium tuberculosis ; classification ; genetics ; isolation & purification ; Ribotyping