The detection of enterotoxins and toxic shock syndrome toxin genes in Staphylococcus aureus by polymerase chain reaction

被引:147
作者
McLauchlin, J
Narayanan, GL
Mithani, V
O'Neill, G
机构
[1] Cent Publ Hlth Lab, PHLS Div Gastrointestinal Infect, Food Safety Microbiol Lab, London NW9 5HT, England
[2] Cent Publ Hlth Lab, Lab Hosp Infect, London NW9 5HT, England
关键词
D O I
10.4315/0362-028X-63.4.479
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
A simple polymerase chain reaction (PCR)-based procedure was developed for the detection of fragments of staphylococcal enterotoxins (SEs) SEA, SEB, SEC, SED, SEE, SEG, SEH, and SEI together with the toxic shock syndrome toxin (TSST-1) genes of Staphylococcus aureus. One hundred and twenty-nine cultures of S. aureus were selected, 39 of which were recovered from 38 suspected staphylococcal food-poisoning incidents. The method was reproducible, and 32 different toxin genotypes were recognized. The presence of SE genes was associated with S. aureus strains reacting with phages in group III, and the TSST-1 gene with phages in group I. There was a 96% agreement between the PCR results for detection of SEA-D and TSST-1 as compared with a commercial reverse passive latex agglutination assay for the detection of SEs from cultures grown in vitro. Enterotoxin gene fragments were detected in S. aureus cultures recovered from 32 of the 38 suspected staphylococcal food poisoning incidents, and of these, 17 were associated with SEE, SEG, SEH, and SEI in the absence of SEA-D. Simple PCR procedures were also developed for the detection of SE directly in spiked food samples, and this was most successfully achieved in mushroom soup and ham. Detection was less successful in three types of cheese and in cream. SEA or SEB were detected by enzyme-linked immunosorbent assay in three food samples (two of which were associated with food poisoning incidents) naturally heavily contaminated with S. aureus: the appropriate SEA or SEB gene fragments were detected directly in these three foods by PCR.
引用
收藏
页码:479 / 488
页数:10
相关论文
共 23 条
[1]  
Barrow GI., 1993, COWAN STEELS MANUAL, V3th
[2]   Rapid and specific detection of toxigenic Staphylococcus aureus:: Use of two multiplex PCR enzyme immunoassays for amplification and hybridization of staphylococcal enterotoxin genes, exfoliative toxin genes, and toxic shock syndrome toxin 1 gene [J].
Becker, K ;
Roth, R ;
Peters, G .
JOURNAL OF CLINICAL MICROBIOLOGY, 1998, 36 (09) :2548-2553
[3]  
BERGDOLL MS, 1981, LANCET, V1, P1017
[4]   RAPID AND SIMPLE METHOD FOR PURIFICATION OF NUCLEIC-ACIDS [J].
BOOM, R ;
SOL, CJA ;
SALIMANS, MMM ;
JANSEN, CL ;
WERTHEIMVANDILLEN, PME ;
VANDERNOORDAA, J .
JOURNAL OF CLINICAL MICROBIOLOGY, 1990, 28 (03) :495-503
[5]  
DESAXE M, 1982, SOC APPLIED BACTERIO, V17, P173
[6]  
Jablonski L. M., 1997, FOOD MICROBIOLOGY FU, P353
[7]   DETECTION OF GENES FOR ENTEROTOXINS, EXFOLIATIVE TOXINS, AND TOXIC SHOCK SYNDROME TOXIN-1 IN STAPHYLOCOCCUS-AUREUS BY THE POLYMERASE CHAIN-REACTION [J].
JOHNSON, WM ;
TYLER, SD ;
EWAN, EP ;
ASHTON, FE ;
POLLARD, DR ;
ROZEE, KR .
JOURNAL OF CLINICAL MICROBIOLOGY, 1991, 29 (03) :426-430
[8]   METACHROMATIC AGAR-DIFFUSION METHODS FOR DETECTING STAPHYLOCOCCAL NUCLEASE ACTIVITY [J].
LACHICA, RUF ;
GENIGEORGIS, C ;
HOEPRICH, PD .
APPLIED MICROBIOLOGY, 1971, 21 (04) :585-+
[9]   ENTEROTOXINS AND TOXIC-SHOCK SYNDROME TOXIN-1 IN NONENTERIC STAPHYLOCOCCAL DISEASE [J].
MARPLES, RR ;
WIENEKE, AA .
EPIDEMIOLOGY AND INFECTION, 1993, 110 (03) :477-488
[10]  
MORRIS CA, 1972, LANCET, V2, P1375