Establishment and application of isothermal multiple-self-matching-initiated amplification (IMSA) in detecting Type II heat-labile enterotoxin of Escherichia coli

• Published in: PloS one Vol. 14; no. 5; p. e0216272
• Main Authors: Liu, Wenxin, Yuan, Chaowen, Zhang, Liguo, Feng, Yufei
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 02.05.2019; Public Library of Science (PLoS)
• Subjects: Analysis; Animal husbandry; Bacteria; Bacterial Toxins - genetics; Beef cattle; Biology and Life Sciences; Child; Child, Preschool; Children; Coliforms; Deoxyribonucleic acid; Diarrhea; Diarrhea - diagnosis; Diarrhea - genetics; Diarrhea - microbiology; DNA; DNA polymerase; Dosage and administration; E coli; EDTA; Enterotoxigenic Escherichia coli - genetics; Enterotoxigenic Escherichia coli - isolation & purification; Enterotoxigenic Escherichia coli - pathogenicity; Enterotoxins; Enzymes; Escherichia coli; Female; Gene amplification; Genes; Health risks; Health screening; Heat-labile enterotoxin; Humans; Male; Matching; Medicine and Health Sciences; Methods; Nucleic Acid Amplification Techniques; Physical Sciences; Public health; Real time; Research and Analysis Methods; Sensitivity analysis; Technology; Travelers; United States > US; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0216272

Enterotoxigenic Escherichia coli (ETEC) constitutes a major cause of diarrhea in young children and animals, particularly in poor regions of the world, as well the traveler's diarrhea in adult individuals. Type II heat-labile enterotoxin (LT-II) from ETEC can cause profuse watery diarrhea, posing a potential threat to public health and animal husbandry. In the present study, isothermal multiple-self-matching-initiated amplification (IMSA) was established to rapidly detect LT-II producing ETEC. The specificity and sensitivity were assessed, and clinical samples were tested. The established IMSA method had good specificity for the detection of LT-II gene with a limit of detection of 25 CFU/mL, i.e. 2 times higher than that of real-time PCR and other two isothermal amplifications (loop-mediated isothermal amplification, LAMP and cross-primer isothermal amplification, CPA). Meanwhile, in 103 clinical Escherichia coli strains isolated from diarrhea samples, 9 strains with LT-II+ gene were detected (8.73%), corroborating real-time PCR, LAMP and CPA data. Therefore, the IMSA technology applied for the detection of LT-II producing ETEC has a good application prospect for screening clinical samples in primary medical units or common laboratories.