Detection of gyrA mutation among clinical isolates of Campylobacter jejuni isolated in Egypt by MAMA PCR

Autores: Said Mayar M, El-Mohamady Hanan, El-Beih Fawkia M, Rockabrand David M, Ismail Tharwat F, Monteville Marshall R, Ahmed Salwa F, et al

Resumen

Introduction: Campylobacter spp are the major cause of enteritis in humans and more than 90% of reported infections are caused by Campylobacter jejuni. Fluoroquinolones such as ciprofloxacin are the antibiotics of choice for treatment. An increase in the frequency of ciprofloxacin-resistant Campylobacter has been reported globally due to a single base mutation (C-257 to T) in codon 86 of the quinolone resistance determining region (QRDR) of the gyrA gene altering the amino acid sequence fron threonine at position 86 to isoleucine (Thr-86 to Ile). Methodology: Campylobacter spp (n = 118) were selected fron a collection of Egyptian isolates spanning 1998 to 2005. The presence of C. jejuni gyrA gene was confirmed in each isolate by a PCR assay amplifying 368bp portion of the gyrA gene. C to T alteration was detected by the mismatch amplification mutation assay MAMA PCR. The MIC of nalidixic acid (NA) and ciprofloxacin (CIP) was determined by E-test. Results: C. jejuni gyrA gene was detected in 100 of the Campylobacter spp studied; the other 18 isolates were found to be Campylobacter coli by lpxA PCR. The mutation was detected in 89 C. jejuni resistant isolates with MIC values (NA; 8 - >256μg/ml) and (CIP; 4 - >32μg/ml). The other 11 sensitive C. jejuni isolates with MIC values (NA; 0.38 – 3μg/ml) and (CIP; 0.03 – 0.125μg/ml) were not amplified by the MAMA primers. There was 100% congruence with MAMA PCR, MIC results and gyrA gene sequence analysis. Conclusions: In Egypt the main mechanism for resistance to fluoroquinolones is an alteration in the gyrA QRDR. MAMA PCR provides an economical and rapid means for screening fluoroquinolone resistance.

Palabras clave: Fluoroquinolone resistance mismatch sequence detection Africa.

2011-04-19   |   261 visitas   |   Evalua este artículo 0 valoraciones

Vol. 4 Núm.9. Septiembre 2010 Pags. 546-554. J Infect Developing Countries 2010; 4(9)