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
| Autor: | Jiann-Hwa Chen, Chien-Shun Chiou, Jeng-Yuan Hsu, Wei-Chang Huang, Jang-Jih Lu, Shiau-Ting Hu, Chen-Cheng Huang, Gwan-Han Shen |
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| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Microbiology (medical)
Tuberculosis lcsh:QR1-502 Microbiology Sensitivity and Specificity lcsh:Microbiology Mycobacterium Mycobacterium tuberculosis Capillary electrophoresis Bacterial Proteins Multiplex polymerase chain reaction medicine Humans hsp65 restriction fragment length polymorphism analysis Capillary electrophoresis Bacteriological Techniques biology Electrophoresis Capillary Chaperonin 60 DNA-Directed RNA Polymerases rpoB duplex polymerase chain reaction rpoB medicine.disease biology.organism_classification bacterial infections and mycoses Molecular biology Parasitology Molecular Diagnostic Techniques Restriction fragment length polymorphism Multiplex Polymerase Chain Reaction circulatory and respiratory physiology Research Article |
| Zdroj: | BMC Microbiology, Vol 12, Iss 1, p 137 (2012) BMC Microbiology |
| ISSN: | 1471-2180 |
| Popis: | 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 rpo B 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 rpo B 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 rpo B 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. |
| Databáze: | OpenAIRE |
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