Detection of vancomycin-resistant enterococci in fecal samples by PCR

被引:94
作者
Satake, S
Clark, N
Rimland, D
Nolte, FS
Tenover, FC
机构
[1] CTR DIS CONTROL & PREVENT, NOSOCOMIAL PATHOGENS LAB BRANCH G08, HOSP INFECT PROGRAM, ATLANTA, GA 30333 USA
[2] VET AFFAIRS MED CTR, MED SERV, ATLANTA, GA 30323 USA
[3] EMORY UNIV HOSP, DEPT PATHOL, ATLANTA, GA 30322 USA
关键词
D O I
10.1128/JCM.35.9.2325-2330.1997
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Surveillance cultures for vancomycin-resistant enterococci (VRE) are time-consuming and expensive for the laboratory to perform. Therefore, we investigated the use of PCR as an alternative method of detecting and identifying VRE directly in fecal samples. PCR primers directed to vanA, vanB, vanC1, vanC2, and enterococcal ligase genes were used to detect and identify VRE in fecal material obtained by rectal or perirectal swabbing, Although PCR-inhibitory substances were present in DNA prepared directly from the swabs, the inhibitory substances could be reduced by processing the nucleic acid with two commercially available DNA preparation columns, Fecal material from 333 swabs was cultured on several selective agar media before and after broth enrichment. DNA was extracted from the fecal material and was analyzed by PCR, By using all four primer sets, only 59 (67.8%) of the samples were positive for vanA. However, after retesting the negative samples with only the vanA primer set, 77 (88.5%) of 87 specimens that were culture positive for Enterococcus faecium containing vanA were positive by PCR, One specimen was PCR positive for the vanA gene but culture negative for enterococci, The specificity of the vanA assay was 99.6%. PCR analysis of enrichment broth samples with all four primers sets after 15 to 18 h of incubation detected 74 (85.1%) of the 87 culture-positive specimens, The specificity of the vanA assay after the enrichment step was 100%, No vanB-containing enterococci were recovered by culture. Since 16 samples can be tested by PCR in 4 h (including electrophoresis), identification of VRE is possible within 8 h of specimen submission at a cost of approximately $10.12/assay. Thus, PCR may be a cost-effective alternative to culture for surveillance of VRE in some hospitals.
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收藏
页码:2325 / 2330
页数:6
相关论文
共 31 条
[1]  
[Anonymous], 1995, INFECT CONT HOSP EP, V16, P105
[2]   GENETICS AND MECHANISMS OF GLYCOPEPTIDE RESISTANCE IN ENTEROCOCCI [J].
ARTHUR, M ;
COURVALIN, P .
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 1993, 37 (08) :1563-1571
[3]  
Centers for Disease Control and Prevention (CDC), 1993, MMWR Morb Mortal Wkly Rep, V42, P597
[4]   CHARACTERIZATION OF GLYCOPEPTIDE-RESISTANT ENTEROCOCCI FROM UNITED-STATES HOSPITALS [J].
CLARK, NC ;
COOKSEY, RC ;
HILL, BC ;
SWENSON, JM ;
TENOVER, FC .
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 1993, 37 (11) :2311-2317
[5]   EMERGENCE OF HIGH-LEVEL RESISTANCE TO GLYCOPEPTIDES IN ENTEROCOCCUS-GALLINARUM AND ENTEROCOCCUS-CASSELIFLAVUS [J].
DUTKAMALEN, S ;
BLAIMONT, B ;
WAUTERS, G ;
COURVALIN, P .
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 1994, 38 (07) :1675-1677
[6]   DETECTION OF GLYCOPEPTIDE RESISTANCE GENOTYPES AND IDENTIFICATION TO THE SPECIES LEVEL OF CLINICALLY RELEVANT ENTEROCOCCI BY PCR [J].
DUTKAMALEN, S ;
EVERS, S ;
COURVALIN, P .
JOURNAL OF CLINICAL MICROBIOLOGY, 1995, 33 (01) :24-27
[7]   RAPID DETECTION OF VANCOMYCIN-RESISTANT ENTEROCOCCI [J].
EDBERG, SC ;
HARDALO, CJ ;
KONTNICK, C ;
CAMPBELL, S .
JOURNAL OF CLINICAL MICROBIOLOGY, 1994, 32 (09) :2182-2184
[8]   AN OVERVIEW OF NOSOCOMIAL INFECTIONS, INCLUDING THE ROLE OF THE MICROBIOLOGY LABORATORY [J].
EMORI, TG ;
GAYNES, RP .
CLINICAL MICROBIOLOGY REVIEWS, 1993, 6 (04) :428-442
[9]   IDENTIFICATION, CLASSIFICATION, AND CLINICAL RELEVANCE OF CATALASE-NEGATIVE, GRAM-POSITIVE COCCI, EXCLUDING THE STREPTOCOCCI AND ENTEROCOCCI [J].
FACKLAM, R ;
ELLIOTT, JA .
CLINICAL MICROBIOLOGY REVIEWS, 1995, 8 (04) :479-&
[10]  
Facklam Richard R., 1995, P308