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Direct detection of Campylobacter jejuni in human stool samples by real-time PCR

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Abstract:

Our purpose was to establish a quick and accurate real-time PCR (rtPCR) method to detect Campylobacter jejuni directly from human diarrheal stool as an alternative to traditional culture methods. To determine the consistency of rtPCR and culture method, 256 clinical diarrheal stool samples and 50 normal stool samples from healthy individuals were examined, and the whole process was double-blinded. Our data showed that the sensitivity of rtPCR in pure cultures and stool was 102 CFU·mL-1 and 103 CFU·g-1, respectively. Of the 256 diarrheal samples, 10 specimens were successfully detected by both methods, whereas two specimens were PCR positive but culture negative. No positive results were found by these two methods in 50 normal specimens. Our data suggested that rtPCR was convenient in operation and time-saving (turnaround time 3.5-4 h), so it could be used for clinical diagnostic and epidemiological purposes.

Notre but était de mettre au point une méthode rapide et fiable de PCR en temps réel (rtPCR) permettant de détecter Campylobacter jejuni directement à partir d’échantillons de selles chez l’humain comparativement aux méthodes de culture traditionnelles. Afin de déterminer le niveau d’uniformité de la rtPCR et de la méthode de culture, 256 échantillons cliniques de selles diarrhéiques et 50 échantillons de selles normales d’individus en bonne santé ont été examinés dans un processus à double-insu. Nos résultats ont démontré que la sensibilité de la rtPCR des cultures pures et des échantillons de selles était de 102 CFU·mL-1et 103 CFU·g-1 respectivement. Des 256 échantillons diarrhéiques, 10 spécimens ont été détectés avec succès par les deux méthodes alors que deux spécimens étaient positifs en PCR mais négatifs en culture. Aucun résultat positif n’a été obtenu par ces deux méthodes à partir des 50 échantillons normaux. Nos résultats ont suggéré que la rtPCR était pratique et rapide (temps d’exécution de 3,5-4 h) de telle sorte qu’elle pourrait être utilisée à des fins de diagnostic clinique ou des fins épidémiologiques.

Document Type: Research Article

Publication date: 2008-09-01

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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