The Study Of Nontuberculous Mycobacteria In The Suspected Pulmonary Tuberculosis Patients In Zhejiang

This study was divided into three parts.Part A:The study of nontuberculous mycobacteria in the suspected pulmonary tuberculosis patients in ZhejiangBackgroundNontuberculous mycobacteria (NTM) belongs to the mycobacterium which is acid-fast staining positive and has gained more and more attention recently. For the research worldly showed that the prevalence of NTM was increasing; and as the pathogens widely existing in the nature environment, NTM could cause serious infections in the immunocompromised patients. The infections could involve multifocal of the patients, such as pulmonary, skin, soft tissue and bone marrow, and were often misdiagnosed as tuberculosis. Our team started to collect the mycobacteria isolates cultured from the sputum of the suspected pulmonary tuberculosis patients since 2011, and until the end of 2013 we had collected 1953 acid-fast straining positive strains from 1831 patients, and identified 113 strains NTM in Demonstration Zone of the prevention and treatment of infectious diseases in Zhejiang Province. We performed this study based on these NTM strains and patients.ObjectivesThis study was performed in Zhejiang Province, southeastern China, to investigate the prevalence, antibiotic susceptibility, and associated clinical, risk and socio-economic factors of NTM pulmonary infections in suspected tuberculosis individuals.MethodologyFrom the local mycobacterial surveillance system,113 NTM isolates were collected from 100 patients with suspected pulmonary tuberculosis between 2011 and 2013. Isolates were identified using mycobacterium identification microarrays and gene sequencing, and in vitro microdilution drug susceptibility testing was subsequently performed.ResultsThe NTM isolation rate was 5.0% in 2011,6.1% in 2012, and 6.2% in 2013 (p>0.05). The main species identified were M. intracellulare, M. avium, M. kansasii and M. abscessus. Drug susceptibility testing confirmed high rates of resistance among M. abscessus strains, while all M. kansasii strains were susceptible to rifampicin, and all M. intracellulare and M. avium strains were susceptible to clarithromycin.ConclusionsM. intracellulare, M. avium, M. kansasii and M. abscessus were the four main species of mycobacteria we found in Zhejiang. In addition, the lower middle-income aged male, the young male manual transient workers and the aged female with the comorbidity of bronchiectasis or pulmonary emphysema were three distinct patient groups. For the high rates of drug resistant M. abscessus isolated from the patients in Zhejiang, the species identification test and in vitro minimum inhibitory concentrations of drugs tested should be used before anti-NTM therapy.Part B:Analysis of the ERIC-PCR genetic fingerprinting map of the mycobacterium intracellulare from ZhejiangBackgroundERIC-PCR (Enterbacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction) could amplify the highly conservative repetitive intergenic sequences from the bacteria, and the genetic fingerprinting map of these products could be used for the homology relationships through the clustering analysis.ObjectivesCurrently it lacks the direct evidence that the mycobacterium intracellulare (M. intracellulare) could infect and spread from man to man. We hold the hyperthosis that the M. intracellulare in Zhejiang had low homology relationships, and analysed the 38 isolates of M. intracellulare by ERIC-PCR.Methodology37 isolates from 36 patients in Zhejiang and 1 standard strain were analysed by ERIC-PCR from 2011 to 2012. Clustering analysis was performed for the homology analysis.ResultsAll the strains had ERIC-PCR products, the sizes were between 100bp to 2000bp, the numbers of bands were between 3 to 12. All the gene finger print types were different, no bands have the same finger type.ConclusionsERIC-PCR could be used for the analysis of molecular typing and homology test of M. intracellulare. The homology of the M. intracellulare from patients in Zhejiang was low.Part C:A case report and study of a multifocal osteomyelitis patient infected by multiple drug resistant nontuberculous mycobacteriaNontuberculous mycobacterium (NTM) has gained increasing attention in recent years for its ability to cause serious complications in immunocompromised patients, and it could involve multifocal of the patients, such as bone marrow, skin, and lymph nodes. But these pathogen lack the attention for a long time because of its low prevalence.Patients of diabetes mellitus are immunocompromised, and osteomyelitis caused by infection is one complication of clinical significance in these patients. Although osteomyelitis caused by nontuberculous mycobacterium infections may have serious consequences for diabetes patients, but few cases have been reported in the world literature. In this case, we report a 69-year-old male patient with diabetes mellitus from Zhejiang, he was infected by Mycobacterium intracellulare from soft tissue to bone marrow. Despite the resistance of the bacilli, the combined antimycobacterial chemotherapy was still able to control the infection. We also provide a global review of the literature in English of osteomyelitis caused by mycobacteria in diabetes mellitus patients.