Rapid and sensitive detection of Plesiomonas shigelloides by loop-mediated isothermal amplification of the hugA gene.
Plesiomonas shigelloides is one of the causative agents of human gastroenteritis, with increasing number of reports describing such infections in recent years. In this study, the hugA gene was chosen as the target to design loop-mediated isothermal amplification (LAMP) assays for the rapid, specific, and sensitive detection of P. shigelloides. The performance of the assay with reference plasmids and spiked human stools as samples was evaluated and compared with those of quantitative PCR (qPCR). No false-positive results were observed for the 32 non-P. shigelloides strains used to evaluate assay specificity. The limit of detection for P. shigelloides was approximately 20 copies per reaction in reference plasmids and 5×10(3) CFU per gram in spiked human stool, which were more sensitive than the results of qPCR. When applied in human stool samples spiked with 2 low levels of P. shigelloides, the LAMP assays achieved accurate detection after 6-h enrichment. In conclusion, the LAMP assay developed in this study is a valuable method for rapid, cost-effective, and simple detection of P. shigelloides in basic clinical and field laboratories in the rural areas of China.
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changping, Beijing, People's Republic of China.
Shuang
S
Meng
Jianguo
J
Xu
Yanwen
Y
Xiong
Changyun
C
Ye
D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D015169:Colony Count, Microbial; D017931:DNA Primers; D005243:Feces / Q000382:microbiology; D005798:Genes, Bacterial; D057230:Limit of Detection; D016981:Plesiomonas / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results
D001426:Bacterial Proteins; D017931:DNA Primers