Development of multiplex oligonucleotide ligation-PCR-universal DNA microarrays for detection of foodborne pathogens

To develop a multiplex oligonucleotide ligation-polymerase chain reaction( MOL-PCR) based universal microarray assay for multiplexed detection of foodborne pathogens. Eight common foodborne pathogens causing bacterial food poisoning were selected as detection models. An upstream and downstream adjac...

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Vydané v:Wei sheng yan jiu Ročník 46; číslo 2; s. 225
Hlavní autori: Wang, Xiaoqiang, Ying, Sisi, Han, Ruijun, Yuan, Jun
Médium: Journal Article
Jazyk:Chinese
Vydavateľské údaje: China 01.03.2017
Predmet:
DNA Primers
DNA, Bacterial
Food Microbiology - instrumentation
Food Microbiology - methods
Foodborne Diseases - genetics
Foodborne Diseases - microbiology
Humans
Multiplex Polymerase Chain Reaction - methods
Oligonucleotide Array Sequence Analysis - instrumentation
Oligonucleotide Array Sequence Analysis - methods
Oligonucleotides
Sensitivity and Specificity
foodborne pathogens
universal DNA microarrays
multiplex oligonucleotide ligation-polymerase chain reaction(MOL-PCR)
ISSN:1000-8020
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Shrnutí:To develop a multiplex oligonucleotide ligation-polymerase chain reaction( MOL-PCR) based universal microarray assay for multiplexed detection of foodborne pathogens. Eight common foodborne pathogens causing bacterial food poisoning were selected as detection models. An upstream and downstream adjacent detection probes were designed within specific primer pair for each of eight pathogens. Target fragments of the eight pathogens were enriched by multiplex PCR and used as ligation templates. Abundant fluorescently labeled single-stranded amplicons containing anti-tag sequences were gained by multiplex ligase detection reaction and asymmetric PCR labeling with universal primers. The products could be detected by hybridization with corresponding tag sequences immobilized on DNA microarrays. The results indicated that the assay could specifically identify all eight pathogens in single and multiple infections. The detection limits were( 1. 1- 8. 5) × 10~2 CFU /mL of pure bacterial cultures. The microarray results f
Bibliografia:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1000-8020