Combined rpoB duplex PCR and hsp65 PCR restriction fragment length polymorphism with capillary electrophoresis as an effective algorithm for identification of Mycobacterial species from clinical isolates

Details

• Alternative Title:
  BMC Microbiol
• Personal Author:
  Huang, Chen-Cheng ; Chen, Jiann-Hwa ; Hu, Shiau-Ting ; Chiou, Chien-Shun ; Huang, Wei-Chang ; Hsu, Jeng-Yuan ; Lu, Jang-Jih ; Shen, Gwan-Han
• Description:
  Background ; Mycobacteria can be quickly and simply identified by PCR restriction-enzyme analysis (PRA), but misidentification can occur because of similarities in band sizes that are critical for discriminating among species. Capillary electrophoresis can provide computer-aided band discrimination. The aim of this research was to develop an algorithm for identifying mycobacteria by combined rpoB duplex PRA (DPRA) and hsp65 PRA with capillary electrophoresis. ; Results ; Three hundred and seventy-six acid-fast bacillus smear-positive BACTEC cultures, including 200 Mycobacterium tuberculosis complexes (MTC) and 176 non-tuberculous mycobacteria (NTM) were analyzed. With combined hsp65 and rpoB DPRA, the accuracy rate was 100% (200 isolates) for the MTC and 91.4% (161 isolates) for the NTM. Among the discordant results (8.6%) for the NTM, one isolate of Mycobacterial species and an isolate of M. flavescens were found as new sub-types in hsp65 PRA. ; Conclusions ; This effective and novel identification algorithm using combined rpoB DPRA and hsp65 PRA with capillary electrophoresis can rapidly identify mycobacteria and find new sub-types in hsp65 PRA. In addition, it is complementary to 16 S rDNA sequencing.
• Subjects:
  • Bacterial Proteins
  • Bacteriological Techniques
  • Chaperonin 60
  • DNA-Directed RNA Polymerases
  • Electrophoresis, Capillary
  • Humans
  • Molecular Diagnostic Techniques
  • Multiplex Polymerase Chain Reaction
  • Mycobacterium
  • Sensitivity and Specificity
  More +
• Source:
  BMC Microbiol. 2012; 12:137.
• Document Type:
  Journal Article
• Volume:
  12
• Download URL:
  https://stacks.cdc.gov/view/cdc/11040/cdc_11040_DS1.pdf
• File Type:
  [PDF - 254.71 KB]
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:aef2feef4f616a90585655fe869866f6b7782f0d59e852e15980b67761985fde