Objective:To determine the accuracy of TB diagnosis of TB in Zambia in the era of increasing HIV prevalence.Methods:Sputum of the clinically diagnosed TB cases was additionally subjected to liquid culture and molecula...Objective:To determine the accuracy of TB diagnosis of TB in Zambia in the era of increasing HIV prevalence.Methods:Sputum of the clinically diagnosed TB cases was additionally subjected to liquid culture and molecular identification.This study distinguished between TB cases confirmed by positive Mycobacterium tuberculosis(M.tuberculosis) cultures and mycobacterial disease caused by non-tuberculous mycobacteria(NTM).Results:Only 49% of the 173 presumptively diagnosed TB cases was M.tuberculosis cultured,while in 13% (22) cases,a combination of M.tuberculosis and NTM was found.In 18% of the patients only NTM were cultured.In 28% ,no mycobacteria was cultivable.HIV positive status was correlated with the isolation of NTM(P【0.05).Conclusions:The diagnosis of tuberculosis based on symptoms, sputum smear and/or chest X-ray leads to significant numbers of false-positive TB cases in Zambia,most likely due to the increased prevalence of HIV.The role of NTM in tuberculosislike disease also seems relevant to the false diagnosis of TB in Zambia.展开更多
We report two cases of pulmonary sequestration infected with nontuberculous mycobacteria(NTM): Mycobacterium avium and Mycobacterium abscessus.Chest computed tomography showed pneumonic consolidation in the right lowe...We report two cases of pulmonary sequestration infected with nontuberculous mycobacteria(NTM): Mycobacterium avium and Mycobacterium abscessus.Chest computed tomography showed pneumonic consolidation in the right lower lobe,which received a systemic blood supply from the descending aorta in both patients.Video-assisted thoracoscopic surgeries were successfully performed and pathologica]examinations revealed multiple caseating granulomas.A review of the literature revealed only seven previous case reports of pulmonary sequestration infected with NTM,and no case with Mycobacterium abscessus has been reported.展开更多
By employing the pUC19 as a backbone,the regulatory and signal sequences which encode kanamycin resistance, and mycobacterial plasmid origin of replication (oriM) were cloned into the pUC19. The recombinant E. Coli-my...By employing the pUC19 as a backbone,the regulatory and signal sequences which encode kanamycin resistance, and mycobacterial plasmid origin of replication (oriM) were cloned into the pUC19. The recombinant E. Coli-mycobacteria shuttle expression plasmid PBCG-8000 was constructed. The PBCG-8000 was able to replicate in both E. Coli and mycobacteria (including BCG) systems, and to confer stable kanamycin resistance upon transformants. The study should facilitate the development of BCG and other mycobacteria into multivalent vaccine vectors.展开更多
Objective Macrolide susceptibility and drug resistance mechanisms of clinical non-tuberculous mycobacteria（NTM） isolates were preliminarily investigated for more accurate diagnosis and treatment of the infection in ...Objective Macrolide susceptibility and drug resistance mechanisms of clinical non-tuberculous mycobacteria（NTM） isolates were preliminarily investigated for more accurate diagnosis and treatment of the infection in China. Methods Four macrolides, including clarithromycin（CLAR）, azithromycin（AZM）, roxithromycin（ROX）, and erythromycin（ERY）, were used to test the drug susceptibility of 310 clinical NTM isolates from six provinces of China with the broth microdilution method. Two resistance mechanisms, 23 S r RNA and erm, were analyzed with nucleotide sequence analysis. Results Varied effectiveness of macrolides and species-specific resistance patterns were observed. Most Mycobacterium abscessus subsp. massiliense were susceptible and all M. fortuitum were highly resistant to macrolides. All the drugs, except for erythromycin, exhibited excellent activities against slow-growing mycobacteria, and drug resistance rates were below 22.2%. Only four highly resistant strains harbored 2,058/2,059 substitutions on rrl and none of other mutations were related to macrolide resistance. G2191 A and T2221 C on rrl were specific for the M. abscessus complex（MABC）. Seven sites, G2140 A, G2210 C, C2217 G, T2238 C, T2322 C, T2404 C, and A2406 G, were specifically carried by M. avium and M. intracellulare. Three sites, A2192 G, T2358 G, and A2636 G, were observed only in M. fortuitum and one site G2152 A was specific for M. gordonae. The genes erm（39） and erm（41） were detected in M. fortuitum and M. abscessus and inducible resistance was observed in relevant sequevar. Conclusion The susceptibility profile of macrolides against NTM was demonstrated. The well-known macrolide resistance mechanisms, 23 S r RNA and erm, failed to account for all resistant NTM isolates, and further studies are warranted to investigate macrolide resistance mechanisms in various NTM species.展开更多
Background: The incidence of mycobacterial infection, in particular M. tuberculosis complex (MTC), is increasing in some Western countries, while nontuberculous mycobacteria (NTM) may be recognized more frequently in ...Background: The incidence of mycobacterial infection, in particular M. tuberculosis complex (MTC), is increasing in some Western countries, while nontuberculous mycobacteria (NTM) may be recognized more frequently in clinical specimens worldwide. The clinical scenario and available histopathology alone are often insufficient to separate these two categories of mycobacterial disease, whose behavior and treatment differ. In particular, NTM may be clinically unsuspected in pathological specimens and the opportunity for culturing missed. Methods: We developed two multiplex PCR assays, which distinguish MTC from NTM by detecting the IS6110 insert in the first tube and discriminating up to 14 NTM reference strains in the second by targeting the 16S-23S rRNA internal transcribed spacer. Test material included 594 routine clinical specimens with diverse pathology;many were granulomas unrelated to mycobacterial infection. About 75% were formalin-fixed paraffin blocks, the remainder mainly cytologic imprints or aspirates on FTA cards submitted on suspicion of mycobacterial infection either to avoid frozen sectioning (with the attendant risk of aerosolisation) or at the time of fine needle aspiration. Results: The paraffinized material yielded 53 MTC positives and the cytological 21 positives. A subset consisting of 337 specimens was also analyzed for NTM and yielded 51 positives. The frequency of simultaneous NTM infection in tuberculous patients was about 17%. Mycobacterium avium complex represented the dominant NTM species overall, showed a predilection for lung and lymph node, and together with M. haemophilum were the second most frequent NTM just behind M. ulcerans/M. marinum in skin and soft tissue, the category displaying the largest NTM diversity. Conclusions: Cytological and deparaffinized tissue analyzed in a new two-tube multiplex PCR allows for specific discrimination of causative agents in mycobacterial infection. MTC is readily distinguished from NTM for appropriate therapy, and NTM presumptively diagnosed at the species level allows appropriate choices of antimicrobials.展开更多
Objective Mycobacterium tuberculosis(Mtb),the causative agent of tuberculosis(TB),causes an estimated 1.6 million human deaths annually,but the pathogenesis of TB remains unclear.Immunity plays a critical role in the ...Objective Mycobacterium tuberculosis(Mtb),the causative agent of tuberculosis(TB),causes an estimated 1.6 million human deaths annually,but the pathogenesis of TB remains unclear.Immunity plays a critical role in the onset and outcome of TB.This study aimed to uncover the roles of innate and adaptive immunity in TB.Methods The gene expression profiles generated by RNA sequencing from human peripheral blood mononuclear cells(PBMCs)stimulated with or without Mtb strain H37Rv antigens were analyzed.A total of 973 differentially expressed mRNAs were identified.Results The differentially expressed genes were enriched in innate immunity signaling functions.The mesenchymal-epithelial transition factor(MET)gene was significantly upregulated in CD14^(+)monocytes.A MET inhibitor improved the uptake of the BCG strain by monocytes and macrophages as well as inhibited the expression of indoleamine 2,3-dioxygenase(IDO).The expression of IDO was increased in PBMCs stimulated with Mtb antigens,and the IDO inhibitor promoted the expression of CD40,CD83,and CD86.Conclusion Our results might provide clues regarding the immunomodulatory mechanisms used by Mtb to evade the host defense system.展开更多
Over the past few decades, there has been a significant increase in the number of mycobacterial species described. Currently, the genus?Mycobacterium?consists of 170 species. Most species are called nontuberculous myc...Over the past few decades, there has been a significant increase in the number of mycobacterial species described. Currently, the genus?Mycobacterium?consists of 170 species. Most species are called nontuberculous mycobacteria (NTM) and are potentially or rarely pathogenic and ubiquitous. One of the main challenges in mycobacteriology is the rapid and precise identification of these microorganisms. In this work, we compared two protein extraction protocols for the identification of 38 reference strains and clinical isolates, representing 27 species, by mass spectrometry (MALDI-TOF MS) to subsequently use the best method for identifying environmental mycobacteria. The results obtained with reference strains and clinical isolates showed that protocol A was effective in identifying 92.1% of mycobacterial specimens at the species level and protocol B, 50%. Therefore, protocol A was evaluated for the rapid identification of 27 environmental mycobacterial isolates. These isolates were subjected to PCR-restriction enzyme analysis (PRA-hsp65). Two isolates were misidentified by PRA-hsp65, whereas MALDI-TOF MS was able to identify them correctly. The results were confirmed by?hsp65 and 16S rRNA gene sequencing. Mass spectrometry has the advantage of being a simpler and faster technique than PRA-hsp65, and our results showed that MALDI-TOF MS is a valuable tool for the identification of environmental mycobacterial isolates.展开更多
We report an unusual manifestation of nontuberculous mycobacterial infection characterized by a giant bursitis on wrist and multiple tenosynovitis with many rice bodies formations. The clinical and radiological examin...We report an unusual manifestation of nontuberculous mycobacterial infection characterized by a giant bursitis on wrist and multiple tenosynovitis with many rice bodies formations. The clinical and radiological examinations are neither rather sensitive nor rather specific. The nuclear imagery of rice bodies formations provides elements of guidance. Cause of absence of the germ isolation, diagnosis was retained on probability items based on a suspicion of arguments beam: clinical, biological, bacteriological and histological. The patient was treated with medical and surgical procedure and provided a satisfactory evolution. At follow-up of 15 months, there were no clinical signs of local recurrence. Through a literature review, the problem of diagnosis of certainty will be discussed.展开更多
Introduction: Mycobacteria pulmonary diseases are chronic illnesses with various impacts on patients’ health status, and wellbeing. These diseases currently represent a global health issue due to increasing burdens a...Introduction: Mycobacteria pulmonary diseases are chronic illnesses with various impacts on patients’ health status, and wellbeing. These diseases currently represent a global health issue due to increasing burdens and the lack of new development on therapeutic options. Policies based on the quality of life may help to improve the management of this chronic respiratory disease;this study was designed to assess the quality of life of patients treated for the pulmonary mycobacterial disease. Materials and Methods: Participants diagnosed with a mycobacterial pulmonary disease were selected from the University Clinical Research Centers’ (UCRC) 2019 mycobacterial cohort database. A telephone interview was conducted using the Medical Outcome Study Short Form (SF-36) which has 36 items evaluating physical and mental wellbeing. Scores range from 0 - 100, with higher scores indicating greater Health-related quality of life (HRQOL). Statistical analysis was performed with SPSS 23.0 and the Fisher test was used to compare percentages. A p-value less than 0.05 was considered significant. Results: A total of 26 participants were reached and interviewed by phone. The mean age was 42 ± 10.6 years, and 76.9% (20/26) were male. The most common cause of the mycobacterial pulmonary disease was Mycobacterium tuberculosis, with 84.6% (22/26). Four cases of Non-Tuberculous Mycobacteria (NTM) were diagnosed including one Mycobacterium avium complex strain (MAC). HIV-positive cases were 46.2% (12/26), and the main respiratory sign was cough for all the participants (100%), followed by dyspnea 46.2% (12/26), chest pain 38.5% (10/26). The mean BMI was 19.7 ± 6.9 kg/m<sup>2</sup>, the mean respiratory rate was 24.7 ± 8.6, and the mean hemoglobin was 11.8 ± 2.2 g/dl. The mean SF-36 score was 75.1 ± 16.6, and impairment was mainly related to mental problems in 59.6%. The mean total score was significantly lower with age more than 42 years than age less than 42 years. But HIV positive and BMI less than 18.5 do not lead to a significant change of mean total score. Conclusion: Patients with mycobacterial pulmonary diseases have more psychological problems than physical ones. Hence the importance of psychological support in their management to improve their quality of life. A large sample size with a deep interview component would be necessary to address limitations in this design.展开更多
文摘Objective:To determine the accuracy of TB diagnosis of TB in Zambia in the era of increasing HIV prevalence.Methods:Sputum of the clinically diagnosed TB cases was additionally subjected to liquid culture and molecular identification.This study distinguished between TB cases confirmed by positive Mycobacterium tuberculosis(M.tuberculosis) cultures and mycobacterial disease caused by non-tuberculous mycobacteria(NTM).Results:Only 49% of the 173 presumptively diagnosed TB cases was M.tuberculosis cultured,while in 13% (22) cases,a combination of M.tuberculosis and NTM was found.In 18% of the patients only NTM were cultured.In 28% ,no mycobacteria was cultivable.HIV positive status was correlated with the isolation of NTM(P【0.05).Conclusions:The diagnosis of tuberculosis based on symptoms, sputum smear and/or chest X-ray leads to significant numbers of false-positive TB cases in Zambia,most likely due to the increased prevalence of HIV.The role of NTM in tuberculosislike disease also seems relevant to the false diagnosis of TB in Zambia.
基金supported by Mid-career Researcher Program through NRF grant funded by the MEST(2011-0015546)
文摘We report two cases of pulmonary sequestration infected with nontuberculous mycobacteria(NTM): Mycobacterium avium and Mycobacterium abscessus.Chest computed tomography showed pneumonic consolidation in the right lower lobe,which received a systemic blood supply from the descending aorta in both patients.Video-assisted thoracoscopic surgeries were successfully performed and pathologica]examinations revealed multiple caseating granulomas.A review of the literature revealed only seven previous case reports of pulmonary sequestration infected with NTM,and no case with Mycobacterium abscessus has been reported.
文摘By employing the pUC19 as a backbone,the regulatory and signal sequences which encode kanamycin resistance, and mycobacterial plasmid origin of replication (oriM) were cloned into the pUC19. The recombinant E. Coli-mycobacteria shuttle expression plasmid PBCG-8000 was constructed. The PBCG-8000 was able to replicate in both E. Coli and mycobacteria (including BCG) systems, and to confer stable kanamycin resistance upon transformants. The study should facilitate the development of BCG and other mycobacteria into multivalent vaccine vectors.
基金supported by the Key Project of the State Key Laboratory for Infectious Disease Prevention and Control[2014SKLID104]the National Key Programs of Mega Infectious Diseases[2013ZX10003002-001]
文摘Objective Macrolide susceptibility and drug resistance mechanisms of clinical non-tuberculous mycobacteria（NTM） isolates were preliminarily investigated for more accurate diagnosis and treatment of the infection in China. Methods Four macrolides, including clarithromycin（CLAR）, azithromycin（AZM）, roxithromycin（ROX）, and erythromycin（ERY）, were used to test the drug susceptibility of 310 clinical NTM isolates from six provinces of China with the broth microdilution method. Two resistance mechanisms, 23 S r RNA and erm, were analyzed with nucleotide sequence analysis. Results Varied effectiveness of macrolides and species-specific resistance patterns were observed. Most Mycobacterium abscessus subsp. massiliense were susceptible and all M. fortuitum were highly resistant to macrolides. All the drugs, except for erythromycin, exhibited excellent activities against slow-growing mycobacteria, and drug resistance rates were below 22.2%. Only four highly resistant strains harbored 2,058/2,059 substitutions on rrl and none of other mutations were related to macrolide resistance. G2191 A and T2221 C on rrl were specific for the M. abscessus complex（MABC）. Seven sites, G2140 A, G2210 C, C2217 G, T2238 C, T2322 C, T2404 C, and A2406 G, were specifically carried by M. avium and M. intracellulare. Three sites, A2192 G, T2358 G, and A2636 G, were observed only in M. fortuitum and one site G2152 A was specific for M. gordonae. The genes erm（39） and erm（41） were detected in M. fortuitum and M. abscessus and inducible resistance was observed in relevant sequevar. Conclusion The susceptibility profile of macrolides against NTM was demonstrated. The well-known macrolide resistance mechanisms, 23 S r RNA and erm, failed to account for all resistant NTM isolates, and further studies are warranted to investigate macrolide resistance mechanisms in various NTM species.
文摘Background: The incidence of mycobacterial infection, in particular M. tuberculosis complex (MTC), is increasing in some Western countries, while nontuberculous mycobacteria (NTM) may be recognized more frequently in clinical specimens worldwide. The clinical scenario and available histopathology alone are often insufficient to separate these two categories of mycobacterial disease, whose behavior and treatment differ. In particular, NTM may be clinically unsuspected in pathological specimens and the opportunity for culturing missed. Methods: We developed two multiplex PCR assays, which distinguish MTC from NTM by detecting the IS6110 insert in the first tube and discriminating up to 14 NTM reference strains in the second by targeting the 16S-23S rRNA internal transcribed spacer. Test material included 594 routine clinical specimens with diverse pathology;many were granulomas unrelated to mycobacterial infection. About 75% were formalin-fixed paraffin blocks, the remainder mainly cytologic imprints or aspirates on FTA cards submitted on suspicion of mycobacterial infection either to avoid frozen sectioning (with the attendant risk of aerosolisation) or at the time of fine needle aspiration. Results: The paraffinized material yielded 53 MTC positives and the cytological 21 positives. A subset consisting of 337 specimens was also analyzed for NTM and yielded 51 positives. The frequency of simultaneous NTM infection in tuberculous patients was about 17%. Mycobacterium avium complex represented the dominant NTM species overall, showed a predilection for lung and lymph node, and together with M. haemophilum were the second most frequent NTM just behind M. ulcerans/M. marinum in skin and soft tissue, the category displaying the largest NTM diversity. Conclusions: Cytological and deparaffinized tissue analyzed in a new two-tube multiplex PCR allows for specific discrimination of causative agents in mycobacterial infection. MTC is readily distinguished from NTM for appropriate therapy, and NTM presumptively diagnosed at the species level allows appropriate choices of antimicrobials.
基金This study was supported by the Thirteen-Fifth Mega-Scientific Project on“Prevention and Treatment of AIDS,Viral Hepatitis and Other Infectious Diseases”(No.2017ZX10201301-007-002)the National Natural Science Foundation of China(No.81571961 and No.82072233)the 309th Hospital(No.2017ZD-007).
文摘Objective Mycobacterium tuberculosis(Mtb),the causative agent of tuberculosis(TB),causes an estimated 1.6 million human deaths annually,but the pathogenesis of TB remains unclear.Immunity plays a critical role in the onset and outcome of TB.This study aimed to uncover the roles of innate and adaptive immunity in TB.Methods The gene expression profiles generated by RNA sequencing from human peripheral blood mononuclear cells(PBMCs)stimulated with or without Mtb strain H37Rv antigens were analyzed.A total of 973 differentially expressed mRNAs were identified.Results The differentially expressed genes were enriched in innate immunity signaling functions.The mesenchymal-epithelial transition factor(MET)gene was significantly upregulated in CD14^(+)monocytes.A MET inhibitor improved the uptake of the BCG strain by monocytes and macrophages as well as inhibited the expression of indoleamine 2,3-dioxygenase(IDO).The expression of IDO was increased in PBMCs stimulated with Mtb antigens,and the IDO inhibitor promoted the expression of CD40,CD83,and CD86.Conclusion Our results might provide clues regarding the immunomodulatory mechanisms used by Mtb to evade the host defense system.
文摘Over the past few decades, there has been a significant increase in the number of mycobacterial species described. Currently, the genus?Mycobacterium?consists of 170 species. Most species are called nontuberculous mycobacteria (NTM) and are potentially or rarely pathogenic and ubiquitous. One of the main challenges in mycobacteriology is the rapid and precise identification of these microorganisms. In this work, we compared two protein extraction protocols for the identification of 38 reference strains and clinical isolates, representing 27 species, by mass spectrometry (MALDI-TOF MS) to subsequently use the best method for identifying environmental mycobacteria. The results obtained with reference strains and clinical isolates showed that protocol A was effective in identifying 92.1% of mycobacterial specimens at the species level and protocol B, 50%. Therefore, protocol A was evaluated for the rapid identification of 27 environmental mycobacterial isolates. These isolates were subjected to PCR-restriction enzyme analysis (PRA-hsp65). Two isolates were misidentified by PRA-hsp65, whereas MALDI-TOF MS was able to identify them correctly. The results were confirmed by?hsp65 and 16S rRNA gene sequencing. Mass spectrometry has the advantage of being a simpler and faster technique than PRA-hsp65, and our results showed that MALDI-TOF MS is a valuable tool for the identification of environmental mycobacterial isolates.
文摘We report an unusual manifestation of nontuberculous mycobacterial infection characterized by a giant bursitis on wrist and multiple tenosynovitis with many rice bodies formations. The clinical and radiological examinations are neither rather sensitive nor rather specific. The nuclear imagery of rice bodies formations provides elements of guidance. Cause of absence of the germ isolation, diagnosis was retained on probability items based on a suspicion of arguments beam: clinical, biological, bacteriological and histological. The patient was treated with medical and surgical procedure and provided a satisfactory evolution. At follow-up of 15 months, there were no clinical signs of local recurrence. Through a literature review, the problem of diagnosis of certainty will be discussed.
文摘Introduction: Mycobacteria pulmonary diseases are chronic illnesses with various impacts on patients’ health status, and wellbeing. These diseases currently represent a global health issue due to increasing burdens and the lack of new development on therapeutic options. Policies based on the quality of life may help to improve the management of this chronic respiratory disease;this study was designed to assess the quality of life of patients treated for the pulmonary mycobacterial disease. Materials and Methods: Participants diagnosed with a mycobacterial pulmonary disease were selected from the University Clinical Research Centers’ (UCRC) 2019 mycobacterial cohort database. A telephone interview was conducted using the Medical Outcome Study Short Form (SF-36) which has 36 items evaluating physical and mental wellbeing. Scores range from 0 - 100, with higher scores indicating greater Health-related quality of life (HRQOL). Statistical analysis was performed with SPSS 23.0 and the Fisher test was used to compare percentages. A p-value less than 0.05 was considered significant. Results: A total of 26 participants were reached and interviewed by phone. The mean age was 42 ± 10.6 years, and 76.9% (20/26) were male. The most common cause of the mycobacterial pulmonary disease was Mycobacterium tuberculosis, with 84.6% (22/26). Four cases of Non-Tuberculous Mycobacteria (NTM) were diagnosed including one Mycobacterium avium complex strain (MAC). HIV-positive cases were 46.2% (12/26), and the main respiratory sign was cough for all the participants (100%), followed by dyspnea 46.2% (12/26), chest pain 38.5% (10/26). The mean BMI was 19.7 ± 6.9 kg/m<sup>2</sup>, the mean respiratory rate was 24.7 ± 8.6, and the mean hemoglobin was 11.8 ± 2.2 g/dl. The mean SF-36 score was 75.1 ± 16.6, and impairment was mainly related to mental problems in 59.6%. The mean total score was significantly lower with age more than 42 years than age less than 42 years. But HIV positive and BMI less than 18.5 do not lead to a significant change of mean total score. Conclusion: Patients with mycobacterial pulmonary diseases have more psychological problems than physical ones. Hence the importance of psychological support in their management to improve their quality of life. A large sample size with a deep interview component would be necessary to address limitations in this design.
