scholarly journals Simultaneous detection of fungal, bacterial, and viral pathogens in insects by multiplex PCR and capillary electrophoresis

2015 ◽  
Vol 30 (2) ◽  
pp. 64-74 ◽  
Author(s):  
Kyu-Won Kwak ◽  
Sung-Hee Nam ◽  
Ji-Young Choi ◽  
Seokhyun Lee ◽  
Hong Geun Kim ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Viral Pathogens

Simultaneous detection of multiple pathogens by multiplex PCR coupled with DNA biochip hybridization

2017 ◽  
Vol 52 (2) ◽  
pp. 186-195 ◽  
Author(s):  
Hsiang-Yun Tung ◽  
Wei-Chen Chen ◽  
Bor-Rung Ou ◽  
Jan-Ying Yeh ◽  
Yeong-Hsiang Cheng ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Clinical Samples ◽  
Single Infection ◽  
Pcr Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Viral Pathogens ◽  
Single Target ◽  
Primer Sets ◽  
Multiple Pathogens

Traditional serological enzyme-linked immunosorbent assay (ELISA) is routinely used to monitor pathogens during quarantine in most animal facilities to prevent possible infection. However, the ELISA platform is a single-target assay, and screening all targeted pathogens is time-consuming and laborious. In this study, to increase sensitivity and to reduce diagnosis time for high-throughput processes, multiplex PCR and DNA biochip techniques were combined to develop a multi-pathogen diagnostic method for use instead of routine ELISA. Eight primer sets were designed for multiplex PCR to detect genes from seven targeted bacterial and viral pathogens. DNA–DNA hybridization was conducted on a biochip following the multiple PCR analysis. Using this method, a total of 24 clinical samples were tested, and the result showed that not only single infection but also co-infection by multi-pathogens can be detected. In conclusion, multiplex PCR coupled with a DNA biochip is an efficient method for detecting multi-pathogens in a reaction. This platform is a useful tool for quarantine services and disease prevention in animal facilities.

Simultaneous Identification of Seven Foodborne Pathogens and Escherichia coli (Pathogenic and Nonpathogenic) Using Capillary Electrophoresis–Based Single-Strand Conformation Polymorphism Coupled with Multiplex PCR

2009 ◽  
Vol 72 (6) ◽  
pp. 1262-1266 ◽  
Author(s):  
MI-HWA OH ◽  
SE-HEE PAEK ◽  
GI WON SHIN ◽  
HAE-YEONG KIM ◽  
GYOO YEOL JUNG ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Capillary Electrophoresis ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Pcr Amplification ◽  
Single Strand Conformation Polymorphism ◽  
Single Strand ◽  
Rrna Gene ◽  
Optimal Separation ◽  
Conformation Polymorphism

The objective of this study was to develop a novel technique for parallel analysis of eight important foodborne microbes using capillary electrophoresis–based single-strand conformation polymorphism (CE-SSCP) coupled with multiplex PCR. Specific primers for multiplex PCR amplification of the 16S rRNA gene were designed, corresponding to eight species of bacteria, including Escherichia coli, Clostridium perfringens, Campylobacter jejuni, Salmonella enterica, Listeria monocytogenes, Vibrio parahaemolyticus, Staphylococcus aureus, and Bacillus cereus, for the species-specific identification and optimal separation of their PCR products in subsequent analysis by CE-SSCP. Multiplex PCR conditions including annealing temperature, extension time, the number of PCR cycles, and primer concentrations were then optimized for simultaneous detection of all target foodborne bacteria. The diagnostic system using CE-SSCP combined with multiplex PCR developed here can be used for rapid investigation of causative agents of foodborne illness. The simplicity and high sensitivity of the method may lead to improved management of safety and illness related to food.

A novel common primer multiplex PCR (CP-M-PCR) method for the simultaneous detection of meat species

Food Control ◽  
2009 ◽  
Vol 20 (4) ◽  
pp. 366-370 ◽  
Author(s):  
Weibin Bai ◽  
Wentao Xu ◽  
Kunlun Huang ◽  
Yanfang Yuan ◽  
Sishuo Cao ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Meat Species ◽  
Pcr Method

Use of a Multiplex PCR System for the Simultaneous Detection of Heat Labile Toxin I and Heat Stable Toxin II Genes of Enterotoxigenic Escherichia coli in Skim Milk and Porcine Stool

1998 ◽  
Vol 61 (2) ◽  
pp. 141-145 ◽  
Author(s):  
HAU-YANG TSEN ◽  
LIANG-ZHAO JIAN ◽  
WAN-RONG CHI
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Skim Milk ◽  
Pcr Primers ◽  
Enterotoxigenic Escherichia Coli ◽  
Farm Animals ◽  
Target Cells ◽  
Heat Stable ◽  
Heat Labile

Enterotoxigenic Escherichia coli (ETEC) strains which produce heat labile and/or heat stable toxins (LT and ST) may cause diarrhea in humans and farm animals. Using PCR primers specific for the LT I and ST II genes, a multiplex PCR system which allows detection of LT I- and ST II-producing ETEC strains was developed. When skim milk was used for a PCR assay, it was found that if target cells in the sample were precultured in MacConkey broth for 8 h prior to PCR as few as 100 cells per ml of the sample could be detected. Without the preculture step, 104 CFU of target cells per 0.2 g of porcine stool specimen were required to generate visible PCR products. The multiplex PCR System can be used for rapid testing of fecal specimens, food and possibly environmental samples for the presence of ETEC strains.

scholarly journals Development of a haemolysin gene-based multiplex PCR for simultaneous detection ofVibrio campbellii,Vibrio harveyiandVibrio parahaemolyticus

2010 ◽  
Vol 50 (2) ◽  
pp. 146-152 ◽  
Author(s):  
S. Haldar ◽  
S.B. Neogi ◽  
K. Kogure ◽  
S. Chatterjee ◽  
N. Chowdhury ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection
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