Detection of gyrA Mutation Among Clinical Isolates of Campylobacter jejuni Isolated in Egypt by MAMA-PCR
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 Campyl...
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The Journal of Infection in Developing Countries
2010-05-01
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| Series: | Journal of Infection in Developing Countries |
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| Online Access: | https://jidc.org/index.php/journal/article/view/963 |
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| author | Mayar Maged Said Hanan El-Mohamady Fawkia M. El-Beih David M. Rockabrand Tharwat F. Ismail Marshall R. Monteville Salwa F. Ahmed John D. Klena Mohamed S. Salama |
| author_facet | Mayar Maged Said Hanan El-Mohamady Fawkia M. El-Beih David M. Rockabrand Tharwat F. Ismail Marshall R. Monteville Salwa F. Ahmed John D. Klena Mohamed S. Salama |
| author_sort | Mayar Maged Said |
| collection | DOAJ |
| description |
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 from threonine at position 86 to isoleucine (Thr-86 to Ile).
Methodology: Campylobacter spp (n = 118) were selected from 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.
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| format | Article |
| id | doaj-art-8aff88ae29cd48f4989e93b2dca7673f |
| institution | DOAJ |
| issn | 1972-2680 |
| language | English |
| publishDate | 2010-05-01 |
| publisher | The Journal of Infection in Developing Countries |
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| series | Journal of Infection in Developing Countries |
| spelling | doaj-art-8aff88ae29cd48f4989e93b2dca7673f2025-08-20T02:57:01ZengThe Journal of Infection in Developing CountriesJournal of Infection in Developing Countries1972-26802010-05-0140910.3855/jidc.963Detection of gyrA Mutation Among Clinical Isolates of Campylobacter jejuni Isolated in Egypt by MAMA-PCRMayar Maged Said0Hanan El-Mohamady1Fawkia M. El-Beih2David M. Rockabrand3Tharwat F. Ismail4Marshall R. Monteville5Salwa F. Ahmed6John D. Klena7Mohamed S. Salama8US. Naval Medical Research Unit#3US. Naval Medical Research Unit#3Ain Shams UniversityUS. Naval Medical Research Unit#3World Health OrganizationUS. NavyUS. Naval Medical Research Unit#3IEIP-CDC (China)Ain Shams University 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 from threonine at position 86 to isoleucine (Thr-86 to Ile). Methodology: Campylobacter spp (n = 118) were selected from 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. https://jidc.org/index.php/journal/article/view/963fluoroquinolone resistancemismatch sequence detectionAfricaMAMA-PCR |
| spellingShingle | Mayar Maged Said Hanan El-Mohamady Fawkia M. El-Beih David M. Rockabrand Tharwat F. Ismail Marshall R. Monteville Salwa F. Ahmed John D. Klena Mohamed S. Salama Detection of gyrA Mutation Among Clinical Isolates of Campylobacter jejuni Isolated in Egypt by MAMA-PCR Journal of Infection in Developing Countries fluoroquinolone resistance mismatch sequence detection Africa MAMA-PCR |
| title | Detection of gyrA Mutation Among Clinical Isolates of Campylobacter jejuni Isolated in Egypt by MAMA-PCR |
| title_full | Detection of gyrA Mutation Among Clinical Isolates of Campylobacter jejuni Isolated in Egypt by MAMA-PCR |
| title_fullStr | Detection of gyrA Mutation Among Clinical Isolates of Campylobacter jejuni Isolated in Egypt by MAMA-PCR |
| title_full_unstemmed | Detection of gyrA Mutation Among Clinical Isolates of Campylobacter jejuni Isolated in Egypt by MAMA-PCR |
| title_short | Detection of gyrA Mutation Among Clinical Isolates of Campylobacter jejuni Isolated in Egypt by MAMA-PCR |
| title_sort | detection of gyra mutation among clinical isolates of campylobacter jejuni isolated in egypt by mama pcr |
| topic | fluoroquinolone resistance mismatch sequence detection Africa MAMA-PCR |
| url | https://jidc.org/index.php/journal/article/view/963 |
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