PMID- 2695745
OWN - NLM
STAT- MEDLINE
DCOM- 19900326
LR  - 20190725
IS  - 0950-382X (Print)
IS  - 0950-382X (Linking)
VI  - 3
IP  - 12
DP  - 1989 Dec
TI  - Multi-gene amplification: simultaneous detection of three virulence genes in 
      diarrhoeal stool.
PG  - 1729-34
AB  - Enterotoxigenic Escherichia coli (ETEC) and Shigella account for a substantial 
      proportion of acute diarrhoeal illnesses among Third-World children. Rapid 
      detection of these infectious agents in faeces followed by the prompt 
      implementation of public health measures could help reduce their spread during 
      the early phase of epidemics. Towards this end, three pairs of synthetic 
      oligonucleotide primers were prepared and shown to hybridize specifically to the 
      genes encoding the heat-stable (ST) and the heat-labile (LT) enterotoxins of ETEC 
      and to invasion-associated loci (ial) of the large Shigella virulence plasmid. 
      When the three primer pairs were used together in the polymerase chain reaction 
      (PCR), the three corresponding genetic loci could be simultaneously amplified 
      using DNA extracted directly from stool; the amplified products were readily 
      detected by ST-, LT- and ial-specific, alkaline phosphatase-labelled 
      oligonucleotide probes (AP probes). The performance of this system was evaluated 
      in a Mayan community in southeastern Mexico, where diarrhoeal illnesses are a 
      common cause of childhood morbidity and mortality. Using only simple and 
      inexpensive laboratory equipment, multigene amplification with these primers and 
      probes led to the identification of ETEC and/or Shigella in the stools of 20 out 
      of 71 children with diarrhoea; the procedure could be completed in seven hours 
      and was more sensitive than conventional diagnostic tests or DNA probes used 
      without amplification.
FAU - Frankel, G
AU  - Frankel G
AD  - Department of Microbiology, Stanford University School of Medicine, California 
      94305.
FAU - Giron, J A
AU  - Giron JA
FAU - Valmassoi, J
AU  - Valmassoi J
FAU - Schoolnik, G K
AU  - Schoolnik GK
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Mol Microbiol
JT  - Molecular microbiology
JID - 8712028
RN  - 0 (DNA Probes)
RN  - 0 (DNA, Bacterial)
SB  - IM
MH  - Base Sequence
MH  - Child, Preschool
MH  - DNA Probes
MH  - DNA, Bacterial/genetics
MH  - Diarrhea/*genetics
MH  - Escherichia coli/*genetics/pathogenicity
MH  - Feces/microbiology
MH  - *Gene Amplification
MH  - Humans
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction
MH  - Shigella/genetics
MH  - Virulence/*genetics
EDAT- 1989/12/01 00:00
MHDA- 1989/12/01 00:01
CRDT- 1989/12/01 00:00
PHST- 1989/12/01 00:00 [pubmed]
PHST- 1989/12/01 00:01 [medline]
PHST- 1989/12/01 00:00 [entrez]
AID - 10.1111/j.1365-2958.1989.tb00158.x [doi]
PST - ppublish
SO  - Mol Microbiol. 1989 Dec;3(12):1729-34. doi: 10.1111/j.1365-2958.1989.tb00158.x.