Isolation, identification and differentiation of Campylobacter spp. using multiplex PCR assay from goats in Khartoum State, Sudan

2017 ◽  
Vol 49 (3) ◽  
pp. 575-581 ◽  
Author(s):  
Atif Elbrissi ◽  
Y. A. Sabeil ◽  
Khalda A. Khalifa ◽  
Khalid Enan ◽  
Osama M. Khair ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Campylobacter Spp

scholarly journals Multiplex PCR Assay for Identifi cation and Differentiation of Campylobacter jejuni and Campylobacter coli Isolates

Folia Medica ◽  
2016 ◽  
Vol 58 (2) ◽  
pp. 95-100 ◽  
Author(s):  
Maria R. Pavlova ◽  
Elina G. Dobreva ◽  
Katucha I. Ivanova ◽  
Galina D. Asseva ◽  
Ivan N. Ivanov ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Campylobacter Jejuni ◽  
Accurate Method ◽  
Clinical Isolates ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
Biochemical Tests ◽  
Multiplex Pcr Assay ◽  
Hydrolysis Of ◽  
Campylobacter Spp

AbstractIntroduction: Campylobacter spp. are important causative agents of gastrointestinal infections in humans. The most frequently isolated strains of this bacterial genus are Campylobacter jejuni and Campylobacter coli. To date, genetic methods for bacterial identification have not been used in Bulgaria. We optimized the multiplex PSR assay to identify Campylobacter spp. and differentiate C. jejuni from C. coli in clinical isolates. We also compared this method with the routinely used biochemical methods.Aim: To identify Campylobacter spp. and discriminate C. coli from C. jejuni in clinical isolates using multiplex PCR assay.Materials and methods: Between February 2014 and January 2015 we studied 93 stool samples taken from patients with diarrheal syndrome and identified 40 species of Campylobacter spp. in them. The clinical material was cultured in microaerophilic atmosphere, the isolated strains being biochemically diff erentiated (hydrolysis of sodium hippurate for C. jejuni, and hydrolysis of indoxyl acetate for C. coli). DNA was isolated from the strains using QiaAmp MiniKit (QIAGEN, Germany). Twenty strains were tested with multiplex PCR for the presence of these genes: cadF, characteristic for Campylobacter spp., hipO for C. jejuni and asp for C. coli.Results and discussion: The biochemical tests identified 16 strains of C. jejuni, 3 strains of C. coli, and 1 strain of C. upsaliensis. After the multiplex PCR assay the capillary gel electrophoresis confirmed 16 strains of C. jejuni, 2 strains of C. coli and 2 strains of Campylobacter spp. - because of the presence of the gene cadF. C. jejuni has the gene hipO, and it is possible that this gene may not be expressed in the biochemical differentiation yielding a negative reaction as a result. In comparison, we can conclude that the genetic differentiation is a more accurate method than the biochemical tests.Conclusion: The multiplex PCR assay is a fast, accurate method for identifi cation of Campylobacter spp. which makes it quite necessary in the clinical diagnostic practice.

scholarly journals A Cytolethal Distending Toxin Gene-Based Multiplex PCR Assay for Detection of Campylobacter spp. in Stool Specimens and Comparison with Culture Method

2012 ◽  
Vol 74 (7) ◽  
pp. 857-862 ◽  
Author(s):  
Sachi SHIRAMARU ◽  
Masahiro ASAKURA ◽  
Haruna INOUE ◽  
Akira NAGITA ◽  
Akio MATSUHISA ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Culture Method ◽  
Toxin Gene ◽  
Cytolethal Distending Toxin ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Stool Specimens ◽  
Campylobacter Spp

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay

2021 ◽  
pp. 106198
Author(s):  
Sunarno ◽  
Khariri ◽  
Fauzul Muna ◽  
Kambang Sariadji ◽  
Yuni Rukminiati ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pcr Assay ◽  
New Approach ◽  
Multiplex Pcr Assay ◽  
Corynebacterium Sp

A multiplex PCR assay to diagnose and quantify Nosema infections in honey bees (Apis mellifera)

2010 ◽  
Vol 105 (2) ◽  
pp. 151-155 ◽  
Author(s):  
Mollah Md. Hamiduzzaman ◽  
Ernesto Guzman-Novoa ◽  
Paul H. Goodwin
Keyword(s):  
Apis Mellifera ◽  
Multiplex Pcr ◽  
Honey Bees ◽  
Pcr Assay ◽  
Multiplex Pcr Assay
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