Analytical comparison of nine PCR primer sets designed to detect the presence of Escherichia coli/Shigella in water samples.

نویسندگان

  • Andrée F Maheux
  • François J Picard
  • Maurice Boissinot
  • Luc Bissonnette
  • Sonia Paradis
  • Michel G Bergeron
چکیده

The analytical performance of 9 different PCR primer sets designed to detect Escherichia coli and Shigella in water has been evaluated in terms of ubiquity, specificity, and analytical detection limit. Of the 9 PCR primer sets tested, only 3 of the 5 primer sets targeting uidA gene and the primer set targeting tuf gene amplified DNA from all E. coli strains tested. However, of those 4 primer sets, only the primer set targeting the tuf gene also amplified DNA from all Shigella strains tested. For the specificity, only the primer sets targeting the uidA gene were 100% specific although the primer sets targeting 16S rRNA, phoE, and tuf genes only amplified Escherichia fergusonii as non-specific target. Finally, the primer set targeting the 16S-ITS-23S gene region, was not specific as it amplified DNA from many other Enterobacteriaceae species. In summary, only the assay targeting the tuf gene detected all E. coli/Shigella strains tested in this study. However, if it becomes important to discriminate between E. coli and E. fergusonii, assays targeting the uidA gene would represent a good choice although none of them were totally ubiquitous to detect of the presence of Shigella strains.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Relationship between Frequency of Escherichia Coli and Prevalence of Salmonella and Shigella Spp. in a Natural River

Introduction: Microbial contamination prediction through detecting the indicator bacteria in natural waters is the first health and environmental step for preventing the transmission of water-associated diseases. This study was designed to determine the correlation between Escherichia coli as the indicator bacterium, on the one hand, and Salmonella and Shigella Spp. As the pathogenic bacteria, ...

متن کامل

Comparison between conventional PCR and PCR - ELISA for detection of Brucella melitensis

Molecular detection techniques are believed to be key tools for both prevention and treatment follow up of brucellosis within live stock and human beings. Consequently rapid, reliable, easy to perform and automated systems for Brucella detection are urgently needed to allow early diagnosis and adequate antibiotic therapy in time. Brucellosis is a worldwide re-emerging zoonosis causing high econ...

متن کامل

Evaluation of ehxA, stx1, and stx2 Virulence Gene Prevalence in Cattle Escherichia coli Isolates by Multiplex PCR

  Today, it is nearly 25 years past from investigation of Shiga toxigenic Escherichia coli (STEC) which is able to produce Shiga toxins and cause different gastroenteritis. Since incidence of gastroenteritis due to STEC is increasing, it's necessary to develop rapid, specific and accurate procedures like PCR. In this study, we used PCR method to detect and identify STEC in cultures of 55 Escher...

متن کامل

A study on detecting and identifying enteric pathogens with PCR.

OBJECTIVE To develop a rapid and definite diagnostic test of bacterial enteritis caused by pathogenic enterobacteria, the most frequent etiologic agent of infectious enteritis in the world. METHODS A set of conventional PCR assays were applied to detect and identify salmonella, shigella, and E. coli O157:H7 directly from pure culture and fecal samples. The general primers of pathogenic entero...

متن کامل

A Comparison between Culture and Multiplex PCR for Detection and Identification of Shigella Species in Patients with Shigellosis from Isfahan Province in 2014-2015

Introduction: Awareness of the species of Shigella has particular importance in the way of dealing with an outbreak, controlling it, and treating patients. The purpose of this study was to use the Multiplex PCR method and comparison with the culture method to detect Shigella species, as well as analysing the antibiotic resistance pattern in its different species. Methods: Simultaneously, the fe...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Water research

دوره 43 12  شماره 

صفحات  -

تاریخ انتشار 2009