Simultaneous Detection of Staphylococcus aureus, Salmonella enterica subsp., Vibrio parahaemolyticus by Multiplex Polymerase Chain Reaction

2010 ◽  
Vol 39 (4) ◽  
pp. 595-601 ◽  
Author(s):  
Yoo-Seok Jeong ◽  
Hee-Kyoung Jung ◽  
Won-Bae Jeon ◽  
Hwa-Jung Seo ◽  
Joo-Heon Hong
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Salmonella Enterica ◽  
Vibrio Parahaemolyticus ◽  
Multiplex Polymerase Chain Reaction ◽  
Simultaneous Detection ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Application Of The Multiplex Polymerase Chain Reaction (M-PCR) For The Screening Of Vibrio Spp. From Rivers In Kuching, Sarawak

1970 ◽  
Vol 5 (2) ◽  
pp. 16-17
Author(s):  
Mickey Vincent ◽  
Lawrance Tuah ◽  
Christy Chan Sien Wei ◽  
Lesley Maurice Bilung ◽  
Kasing Apun
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Parahaemolyticus ◽  
Multiplex Polymerase Chain Reaction ◽  
Vibrio Vulnificus ◽  
Simultaneous Detection ◽  
Chain Reaction ◽  
Single Tube ◽  
Predominant Species ◽  
Polymerase Chain ◽  
Vibrio Spp

The present study was conducted to investigate the occurrence of Vibrio spp. from selected rivers in Kuching,Sarawak (Malaysia) using Multiplex Polymerase Chain Reaction (m-PCR). During the 6-month study period,19 samples were collected monthly from 7 rivers, followed by simultaneous detection of three Vibrio spp.,Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus, in a single tube PCR reaction. Three sets ofprimers targeting the thermolabile (tl), outer membrane protein (ompW) and hemolysin/cytolysin genes(vulCulsl) of V. parahaemolyticus, V. cholerae and V. vulnificus, respectively, were used. The results indicatedthat V. parahaemolyticus was the predominant species, occurring approximately 60.9% throughout thesampling period, followed by V. cholerae (23.1%) and V. vulnificus (16.0%). The months of July andDecember were found to be the months where all three Vibrio spp. were found to be at higher frequencies inthe river samples. Results analyzed also indicated that the rivers with the highest prevalence of the three Vibriospp. were Tambak Sejingkat, followed by Sungai Jernang and Sungai Tabuan. We conclude that m-PCR is apowerful and useful tool for the rapid and simultaneous detection of V. parahaemolyticus, V. cholerae and V.vulnificus from the riverine environments without the need for isolation and culturing. Furthermore, thismethod is highly specific, and could be applied in diagnostic laboratories for larger scale epidemiologicalinvestigations of Vibrio spp.

Development of a Multiplex Polymerase Chain Reaction Assay for Detection and Differentiation of Staphylococcus aureus in Dairy Products†

2001 ◽  
Vol 64 (5) ◽  
pp. 664-668 ◽  
Author(s):  
SUDHIR TAMARAPU ◽  
JOHN L. McKILLIP ◽  
MARYANNE DRAKE
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Dairy Products ◽  
Multiplex Polymerase Chain Reaction ◽  
Skim Milk ◽  
Cheddar Cheese ◽  
Polymorphism Analysis ◽  
Chain Reaction ◽  
Polymerase Chain

A multiplex polymerase chain reaction (PCR) assay was developed for the detection and differentiation of enterotoxigenic Staphylococcus aureus in dairy products. A solvent extraction procedure was successfully modified for extraction of S. aureus DNA from 10 ml of artificially contaminated skim milk or 20 g cheddar cheese. Primers targeting the enterotoxin C gene (entC) and thermostable nuclease gene (nuc) were used in the multiplex PCR. PCR products were confirmed using restriction fragment length polymorphism analysis. DNA was consistently quantified and amplified by uniplex PCR from 10 CFU/ml of S. aureus in skim milk or 10 CFU/20 g cheddar cheese. The sensitivity of the multiplex PCR was 100 CFU/ml of skim milk or 100 CFU/20 g cheddar cheese. The developed methodology allows presumptive identification and differentiation of enterotoxigenic S. aureus in less than 6 h.

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