title: Identification of mycobacterial species by PCR restriction enzyme analysis of the hsp65 gene—an Indian experience.
abstract: Nowadays, nontuberculous mycobacteria (NTM) often cause pulmonary and extrapulmonary disease. Species identification of NTM determines the line of treatment and management of the disease. The routine diagnostic methods, i.e., smear microscopy and biochemical identification, of nontuberculous mycobacteria are tedious and time consuming and not all laboratories can perform these tests on a routine basis. A PCR targeting the hsp65 gene was implemented using standard strains and was applied to 109 clinical isolates. The PCR-amplified product was subjected to restriction enzyme analysis using BstEII and HaeIII. The results obtained were compared with that of biochemical tests. Of 109 NTM, 107 were identified to species level. PCR plus restriction enzyme analysis (PRA) identified 12 types of NTM. Common species identified were Mycobacterium chelonae (32), a rapid growing NTM, and Mycobacterium avium complex (21), among the slow growing NTM. PRA and biochemical identification showed 95.32% (102/107) concordant results. PRA is fast, cheap, and accurate for identification of potentially pathogenic NTM.
authors.affiliation: Department of Microbiology, National Institute of Tuberculosis and Respiratory Diseases, Sri Aurobindo Marg, New Delhi 110030, India.
authors.forename: Ajoy Kumar
authors.initials: AK
authors.lastname: Verma
authors.forename: Gavish
authors.initials: G
authors.lastname: Kumar
authors.forename: Jyoti
authors.initials: J
authors.lastname: Arora
authors.forename: Paras
authors.initials: P
authors.lastname: Singh
authors.forename: Vijay Kumar
authors.initials: VK
authors.lastname: Arora
authors.forename: Vithal Prasad
authors.initials: VP
authors.lastname: Myneedu
authors.forename: Rohit
authors.initials: R
authors.lastname: Sarin
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D018834:Chaperonin 60 / Q000235:genetics*; D006801:Humans; D007194:India; D009165:Mycobacterium Infections, Nontuberculous / Q000382:microbiology*; D009170:Nontuberculous Mycobacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D015183:Restriction Mapping / Q000379:methods*
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; C074119:heat-shock protein 65, Mycobacterium
keywords: analyse par enzyme de restriction; mycobactéries non tuberculeuses; nontuberculous mycobacteria; polymerase chain reaction; restriction enzyme analysis; réaction de la polymérase en chaîne