| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | The selection of eight gene targets (hsp65 631, katG 463, 16S rRNA 1429, gyrA 95, gyrB (675+756), and gyrB (675), gyrB (756), gyrB (1410) were based on the primary information provided in a previous publication (Bouakaze et al., 2010). However, employing the software from GenScript (www. genscript.com/tools.html#biology) new primers were designed for conducting PCR, Real Time PCR and their corresponding minor groove binding (MGB) fluorescent labeled probes and for generating amplicons longer than 300 bp for sequencing experiments for five gene targets hsp65 631, gyrA 95, gyrB (675), gyrB (756) and gyrB (1410) in this study, while retaining the original primers recommended for other three gene targets 16S rRNA 1429, katG 463 and gyrB (756+675) by Bouakaze et al. (2010); the details are provided in Table 1. | Get A Quote |
Accurate identification of Mycobacterium strains is necessary from a diagnostic standpoint. Differences in gyrA 95 and gyrB (1410) genome due to specific single nucleotide polymorphism (SNP) were used to design RealTime PCRs that can discriminate Mycobacterium tuberculosis (MT) from Mycobacterium bovis (MB) belonging to the Mycobacterium Tuberculosis Complex (MTBC). The Real-Time PCR employing the VIC labeled gyrA 95 probe specifically detected the ATCC reference strain of MT H37Rv and eight field strains as MT, but not MB or other members of MTBC and other micro-organism tested. The VIC labeled gyrA 95 assay could detect up to 450 fg of MT. The FAM labeled gyrB (1410) specifically detected MB, but not MT or ot... More
