Use of real time polymerase chain reaction for detection of M. tuberculosis, M. avium and M. kansasii from clinical specimens

BMC Infectious Diseases

Research Article

Enock Matovu¹  Eddie Wampande²  Moses Joloba³  Joseph Akol⁴  Henry Ssenyungule⁴  Arnold Bainomugisa⁴  Sam Ogwang⁴  Chris Muchwa⁴  Paul Mubiri⁵ 

[1] Department of Bio molecular Resources and Bio laboratory Sciences, College of Veterinary Medicine, Animal resource and Bio-security, Kampala, Uganda;Department of Medical Microbiology, College of Health sciences, Makerere University, Kampala, Uganda;Department of Bio molecular Resources and Bio laboratory Sciences, College of Veterinary Medicine, Animal resource and Bio-security, Kampala, Uganda;Department of Medical Microbiology, College of Health sciences, Makerere University, Kampala, Uganda;Uganda-CASE Research Collaboration, Kampala, Uganda;Joint Clinical Research Centre, P.O. Box 10005, Kampala, Lubowa, Uganda;Uganda-CASE Research Collaboration, Kampala, Uganda;Uganda-CASE Research Collaboration, Kampala, Uganda;

关键词: M. tuberculosis;    M. avium;    M. kansasii;    Real time PCR;   

DOI  :  10.1186/s12879-015-0921-0

 received in 2014-12-27, accepted in 2015-04-01,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundThe incidence of M. tuberculosis (MTB) and non tuberculous Mycobacterium species (NTMs) like M. avium and M. kansasii has increased due to Human Immunodeficiency Virus (HIV) epidemic. Therefore accurate, rapid and cost effective methods for the identification of these NTMs and MTB are greatly needed for appropriate TB management. Thus in this study we evaluated the performance of Lightcycler® Mycobacterium detection assay to detect MTB, M. avium and M. kansasii in sputum specimens.MethodsA total of 241 baseline minimally processed sputum specimens from individual adult TB suspected patients were analyzed by Mycobacterium detection assay (Real-time-PCR) on a LightCycler 480® while using liquid culture as a reference standard.ResultsReal time PCR had a sensitivity of 100% (95% CI 96–100) and 100% (CI 19–100) for detection of MTB and M. avium respectively. Additionally the assay had a specificity of 99% (95% CI 96–99) and 95% (95% CI 91–97) for identification of MTB and M. avium respectively. The positive predictive value (PPV) for Real time PCR to identify MTB and M. avium among the specimens was 98% (95% CI 94–99) and 15% (95% CI 2–45) respectively. The kappa statistics for Real time PCR to identify MTB and M. avium was 0.9 (95% CI 0.9–1.0) and 0.3 (95% CI–0.03–0.5) respectively. The median time to detection for Real time PCR assay was 2 hours while overall median time to detection for MGIT-positive cultures was 8 days. The sample unit cost for Real time PCR was $ 12 compared to $ 20 for the reference liquid culture.ConclusionThe Light cycler® Mycobacterium detection assay rapidly and correctly identified MTB and M avium thus has the potential to be adopted in a clinical setting.

【 授权许可】

CC BY   
© Bainomugisa et al.; licensee BioMed Central. 2015