Simultaneous detection of six human diarrheal pathogens by using DNA microarray combined with tyramide signal amplification.
Multiplex PCR and DNA microarray were combined with tyramide signal amplification (TSA) to develop a reliable method suitable for simultaneous detection of six species of human diarrheal pathogens (Yersinia enterocolitica, Shigella spp, Salmonella typhi, Brucella spp, Vibrio cholera and Escherichia coli O157:H7). Meanwhile, our method could distinguish V. cholera serotype O1 from O139, and O157:H7 from O157: non-H7. This assay conferred a specificity of 100% for target pathogens. The limit of detection was 103 degrees CFU/mL approximately. The results of 98.6% (357/362) clinical specimens and 100% (5/5) mocked double-blind samples were the same to that from conventional assay. Consequently this assay is sensitive and a specific tool suitable for diagnostic detection and surveillance of multiple human pathogens.
Jin
Dazhi
D
Beijing Institute of Radiation Medicine, No. 27 Taiping road, Beijing, China. dazhijin_y@hotmail.com
Qi
Hongjuan
H
Chen
Suhong
S
Zeng
Ting
T
Liu
Qiqi
Q
Wang
Shengqi
S
D015342:DNA Probes; D003967:Diarrhea / Q000382:microbiology*; D006090:Gram-Negative Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016905:Gram-Negative Bacterial Infections / Q000382:microbiology; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014439:Tyramine / Q000032:analysis*
D015342:DNA Probes; D014439:Tyramine