Influence of enrichment media and application of a PCR based method to detect Salmonella in poultry industry products and clinical samples

被引:52
作者
Schrank, IS
Mores, MAZ
Costa, JLA
Frazzon, APG
Soncini, R
Schrank, A
Vainstein, MH
Silva, SC [1 ]
机构
[1] Univ Fed Rio Grande do Sul, Ctr Biotechnol, Porto Alegre, RS, Brazil
[2] Univ Fed Rio Grande do Sul, Dept Mol Biol & Biotechnol, Porto Alegre, RS, Brazil
[3] Sadia SA, Cent Lab, BR-89700000 Concordia, SC, Brazil
[4] Univ Fed Rio Grande do Sul, Dept Microbiol, Porto Alegre, RS, Brazil
[5] Univ Fed Rio Grande do Sul, Dept Patol Clin Vet, Porto Alegre, RS, Brazil
关键词
Salmonella enterica; PCR; chicken-bacteria; diagnostic methods;
D O I
10.1016/S0378-1135(01)00350-9
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
To attempt the rapid detection of Salmonella enterica, we have coupled a culture procedure with PCR amplification of the genus-specific invElinvA genes. The method was applied to different kinds of samples from the poultry industry and evaluated by using hydrolyzed feather meal, meat meal, litter and viscera, all experimentally inoculated with a known number of Salmonella followed by cultivation in selenite-cystine broth prior to the PCR reaction. The expected 457 bp specific DNA fragment could be amplified from dilutions containing as few as 5.7 CFU, indicating that the PCR technique can be successfully coupled with culture in an enrichment broth to distinguish Salmonella species from other enteric bacteria present in samples from the poultry industry. Tetrathionate broth proved to be a much better enrichment media compared to selenite-cystine when the presence of Salmonella was evaluated by PCR in 1-day-old chicks experimentally infected with known numbers of Salmonella. Samples included cecal tonsils and viscera, collected at 48 h and 7 days postinfection. The PCR technique was more sensitive in detecting infected animals than the standard microbiological procedure, which detected only 47% of all PCR positive samples. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:45 / 53
页数:9
相关论文
共 20 条
[1]   DETECTION OF SALMONELLA-ENTERITIDIS IN FECES FROM POULTRY USING BOOSTER POLYMERASE CHAIN-REACTION AND OLIGONUCLEOTIDE PRIMERS SPECIFIC FOR ALL MEMBERS OF THE GENUS SALMONELLA [J].
COHEN, ND ;
MCGRUDER, ED ;
NEIBERGS, HL ;
BEHLE, RW ;
WALLIS, DE ;
HARGIS, BM .
POULTRY SCIENCE, 1994, 73 (02) :354-357
[2]   COMPARISON OF THE POLYMERASE CHAIN-REACTION USING GENUS-SPECIFIC OLIGONUCLEOTIDE PRIMERS AND MICROBIOLOGIC CULTURE FOR THE DETECTION OF SALMONELLA IN DRAG-SWABS FROM POULTRY HOUSES [J].
COHEN, ND ;
WALLIS, DE ;
NEIBERGS, HL ;
MCELROY, AP ;
MCGRUDER, ED ;
DELOACH, JR ;
CORRIER, DE ;
HARGIS, BM .
POULTRY SCIENCE, 1994, 73 (08) :1276-1281
[3]   SALMONELLA SEROGROUP-C2 AND SEROGROUP-C3 IDENTIFIED BY AGGLUTINATION USING AN IMMUNOGLOBULIN-G3(K) MONOCLONAL-ANTIBODY (32-1-E3) REACTIVE WITH A SOMATIC FACTOR 8-LIKE POLYSACCHARIDE ANTIGEN [J].
DUFFEY, PS ;
KANI, JC ;
LEE, JO ;
HILL, WJ ;
KOKKA, R .
JOURNAL OF CLINICAL MICROBIOLOGY, 1992, 30 (12) :3050-3057
[4]  
EDWARDS P.R., 1972, IDENTIFICATION ENTER
[5]   SEROLOGY OF SHIGELLA [J].
EWING, WH ;
LINDBERG, AA .
METHODS IN MICROBIOLOGY, 1984, 14 :113-142
[6]   ANALYSIS OF SALMONELLA-ENTERITIDIS ISOLATES BY ARBITRARILY PRIMED PCR [J].
FADL, AA ;
NGUYEN, AV ;
KHAN, MI .
JOURNAL OF CLINICAL MICROBIOLOGY, 1995, 33 (04) :987-989
[7]  
FLUIT AC, 1995, J CLIN MICROBIOL, V33, P1046, DOI 10.1128/JCM.33.4.1046-1047.1995
[8]  
GALAN JE, 1992, J BACTERIOL, V174, P4338
[9]   MOLECULAR DIAGNOSTICS - ISSUES OF UTILIZATION, REGULATION AND ORGANIZATION [J].
GARRETT, CT ;
FERREIRACENTENO, A ;
NASIM, S .
CLINICA CHIMICA ACTA, 1993, 217 (01) :85-103
[10]   PRINCIPLES OF SALMONELLA ISOLATION [J].
HARVEY, RWS ;
PRICE, TH .
JOURNAL OF APPLIED BACTERIOLOGY, 1979, 46 (01) :27-56