Development of a Single-Tube, Cell Lysis-Based, Genus-Specific PCR Method for Rapid Identification of Mycobacteria: Optimization of Cell Lysis, PCR Primers and Conditions, and Restriction Pattern Analysis

Details

• Personal Author:
  Khan IUH ; Yadav JS
• Description:
  A single-tube PCR method was developed for efficient identification of nontuberculous mycobacteria (NTM) and their environmental isolates in about 3 h without conventional DNA isolation. The following three steps were optimized or developed: (i) a simple, 6-min direct cell lysis protocol as a PCR prestep for generation of DNA-template, (ii) an improved Mycobacterium-specific PCR amplification protocol with a broader species specificity using newly designed primers targeting a 228-bp region of the 65-kDa heat shock protein (hsp) gene and optimal PCR amplification conditions, and (iii) a genus-specific restriction analysis of the PCR product for conclusive identification of the unknown NTM isolates. [Description provided by NIOSH]
• Subjects:
  Microbiology Occupational Health Safety
• Keywords:
  Deoxyribonucleic Acids DNA Metalworking Fluids Microbial Test Systems Microorganisms MWFs Mycobacterium
• ISSN:
  0095-1137
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  Ohio ; OSHA Region 5
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  453-457
• Volume:
  42
• Issue:
  1
• NIOSHTIC Number:
  nn:20056251
• Citation:
  J Clin Microbiol 2004 Jan; 42(1):453-457
• Contact Point Address:
  Jagjit S. Yadav, Molecular Toxicology Division, Department of Environmental Health, University of Cincinnati Medical Center, Kettering Laboratory, 3223 Eden Ave., Room 138, Cincinnati, OH 45267-0056
• Email:
  jagjit.yadav@uc.edu
• Federal Fiscal Year:
  2004
• NORA Priority Area:
  Research Tools and Approaches: Exposure Assessment Methods
• Performing Organization:
  University of Cincinnati
• Peer Reviewed:
  True
• Start Date:
  20010930
• Source Full Name:
  Journal of Clinical Microbiology
• End Date:
  20170731
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:ab1fcec577daf646593d2e6b69976e359dc12ff986ee23a7154db475b90ce0a12cc39759c9b72997decd0460082bd05f7606d3bb79c161e7018041276915cc15
• Download URL:
  https://stacks.cdc.gov/view/cdc/213893/cdc_213893_DS1.pdf
• File Type:
  [PDF - 461.68 KB ]