Use of macroporous polypropylene filter to allow identification of bacteria by PCR in human fecal samples

被引:26
作者
Cavallini, A
Notarnicola, P
Berloco, P
Lippolis, A
Di Leo, A
机构
[1] IRCCS S de Bellis, Sci Inst Digest Dis, Biochem Lab, I-70013 Castellana Grotte, BA, Italy
[2] IRCCS S de Bellis, Sci Inst Digest Dis, Clin Pathol Lab, I-70013 Castellana Grotte, BA, Italy
关键词
feces; PCR inhibitors; polypropylene filter; polysaccharides;
D O I
10.1016/S0167-7012(99)00122-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The detection of pathogenic bacteria directly in human fecal specimens by PCR, requires removal of PCR-inhibitory substances. To investigate whether five different macroporous filters (polypropylene, nylon, polyester, polyethylene, fluorocarbon) could retain polysaccharides, major PCR inhibitors, an in vitro model and human fecal samples were used. The in vitro model consisted of Xanthum gum solutions (3 mg/ml PBS), a bacterial polysaccharide, to which Helicobacter pylori cells were added. Fecal samples from healthy volunteers were spiked with H. pylori and Mycobacterium paratuberculosis cells. Polysaccharide concentrations were significantly reduced only by the polypropylene but not by the other filters. Accordingly, both Xanthum gum solutions and spiked fecal specimens became PCR positive only after filtration with the polypropylene filter. We conclude that this filter can be used to prepare a bacterial DNA template suitable for PCR analysis from human feces. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:265 / 270
页数:6
相关论文
共 27 条
[1]   Immunomagnetic separation and PCR detection show shigellae to be common faecal agents in children from urban marginal communities of Costa Rica [J].
Achi, R ;
Mata, L ;
Siles, X ;
Lindberg, AA .
JOURNAL OF INFECTION, 1996, 32 (03) :211-218
[2]  
Ali SA, 1998, BIOTECHNIQUES, V25, P975
[3]  
CHAN JWY, 1995, BIOTECHNIQUES, V18, P419
[4]  
CHOMCZYNSKI P, 1995, BIOTECHNIQUES, V19, P942
[5]   SENSITIVE DETECTION OF HELICOBACTER-PYLORI BY USING POLYMERASE CHAIN-REACTION [J].
CLAYTON, CL ;
KLEANTHOUS, H ;
COATES, PJ ;
MORGAN, DD ;
TABAQCHALI, S .
JOURNAL OF CLINICAL MICROBIOLOGY, 1992, 30 (01) :192-200
[6]  
DEBROT MA, 1997, BIOTECHNIQUES, V23, P616
[7]   Detection of Mycobacterium paratuberculosis in stool samples of patients with inflammatory bowel disease by IS900-based PCR and colorimetric detection of amplified DNA [J].
Del Prete, R ;
Quaranta, M ;
Lippolis, A ;
Giannuzzi, V ;
Mosca, A ;
Jirillo, E ;
Miragliotta, G .
JOURNAL OF MICROBIOLOGICAL METHODS, 1998, 33 (02) :105-114
[8]   A METHOD FOR PREPARATION OF FECAL DNA SUITABLE FOR PCR [J].
DEUTER, R ;
PEITSCH, S ;
HERTEL, S ;
MULLER, O .
NUCLEIC ACIDS RESEARCH, 1995, 23 (18) :3800-3801
[9]   COLORIMETRIC METHOD FOR ROUTINE MEASUREMENT OF DIETARY FIBER AS NONSTARCH POLYSACCHARIDES - A COMPARISON WITH GAS-LIQUID-CHROMATOGRAPHY [J].
ENGLYST, HN ;
HUDSON, GJ .
FOOD CHEMISTRY, 1987, 24 (01) :63-76
[10]   IMMUNOMAGNETIC SEPARATION AND PCR FOR DETECTION OF HELICOBACTER-PYLORI IN WATER AND STOOL SPECIMENS [J].
ENROTH, H ;
ENGSTRAND, L .
JOURNAL OF CLINICAL MICROBIOLOGY, 1995, 33 (08) :2162-2165