Nonradioactive polynucleotide gene probe assay for the detection of three virulence toxin genes in diarrhoeal stools.
The present study describes a nonisotopic DNA-DNA hybridization assay for the detection of enterotoxigenic Escherichia coli (ETEC) in which the probes were labelled with the hapten molecule digoxigenin and after hybridization, DNA hybrids were detected by antidigoxigenin alkaline phosphatase conjugate. A blinded study carried out on a battery of enterotoxigenic and nonenterotoxigenic Esch. coli by dig-probe hybridization assay were compared with the results of a radiolabelled toxin gene probe hybridization assay (performed earlier). The three digoxigenin labelled probes gave a 100 per cent specificity and sensitivities of 95.45, 100 and 100 per cent for LT, STh and STp respectively. These results were comparable to those with the radioactive probes.
Bright
B D
BD
Department of Paediatrics, All India Institute of Medical Sciences, New Delhi.
Kumar
R
R
Jailkhani
B
B
Srivastava
R
R
Bhan
M K
MK
D002648:Child; D015342:DNA Probes / Q000235:genetics; D003967:Diarrhea / Q000382:microbiology*; D004076:Digoxigenin; D004768:Enterotoxins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D004927:Escherichia coli Infections / Q000382:microbiology*; D005243:Feces / Q000737:chemistry / Q000382:microbiology*; D005798:Genes, Bacterial; D006801:Humans; D009693:Nucleic Acid Hybridization; D012680:Sensitivity and Specificity; D014774:Virulence / Q000235:genetics
D015342:DNA Probes; D004768:Enterotoxins; D004076:Digoxigenin