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BMC microbiology [electronic resource]
2012-07-09 03:34:43

Combined

Descrição: 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 the 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 16S rDNA sequencing.

Identificador: doi:10.1186/1471-2180-12-137
Volume: 0
Página: 1 a


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