Detection of Isoniazid-Resistant Mycobacterium tuberculosis Strains by a Multiplex Allele-Specific PCR Assay Targeting katG Codon 315 Variation

Multidrug-resistant tuberculosis: rapid detection of resistance to rifampin and high or low levels of isoniazid in clinical specimens and isolates

“…For the one isolate which had a discrepant result by genotype that was susceptible to RIF, the rpoB gene was subjected to sequencing by a previously described method [16]. Sequencing of the four discrepant isolates and the four isolates obtained from the four discrepant clinical samples was performed in the following regions: katG (codons 315 and 463), fabG1-inhA and the oxyR-ahpC intergenic region using primers previously described [17][18][19]. The PCR conditions used for the katG codon 315, the fabG-inhA and the oxyR-ahpC intergenic region were as follows: primers katg1F and katg4F (katG), 519 and 520 (fabG1-inhA) and tomap1 and tomap2 (oxyRahpC) using AmpliTaq Gold (Applied Biosystems, Foster City, CA) and 1.5 mM MgCl 2 (2.5 mM for oxyR-ahpC), with 30 cycles of 1 min at 95°C, 40 s at 62°C and 1 min at 72°C.…”

Schneller Nachweis von Mutationen im gyrA-Gen von Mycobacterium tuberculosis mit Relevanz für die Entstehung von Medikamentenresistenzen gegen Chinolone mittels der Real-Time PCR auf dem LightCycler® System

“…Exemplarisch sei hier der Nachweis der KatG 315 Mutation aufgrund der RLFP (Nachamkin, Kang et al 1997;Rinder, Feldmann et al 1999) oder mittels einer Allel-spezifischen Multiplex-PCR (Mokrousov, Otten et al 2002) erwähnt. Die Analyse des RRDR erfolgt aufgrund des längeren Genabschnitts meist durch die Hybridisierung von PCR-Produkten mit Oligonucleotiden.…”

The Susceptibility of katG Ser315Thr Mycobacterium tuberculosis mutant Against Anti Tuberculosis Drugs

“…One set of primers katG IF (5'-AGCTCGTATGGCACCGGAAC) and katG 4R (5'-AACGGGTCCGGGATGGTG-3') were used for katG fragment amplifi cation (position 904 to 1103 in H37Rv) (Mokrousov, 2002). Amplified fragments were digested with MspI.…”

“…Mutated codon 315 will be digested by MspI, resulting in 7 fragments ; 14, 6, 21, 6, 11, 10, and 132 bps. Digestion of non mutant katG Ser315Thr resulted in 6 fragments; 14 , 6, 6,11,10, and 153 bps (adapted from Mokrousov et al, 2002). The PCR-RFLP analysis was confi rmed with DNA sequencing analysis.…”

“…This mutation is strongly associated with MDR mutant and transmitted easily among M. tuberculosis strain. Mutation may result in decrease activity of catalase-peroxidase and deletion of this gene may cause loss of the enzyme activity ( Mokrousov, 2002;Xueqiong et al, 2006;Hu et al, 2010).…”