Identification of parasitic nematodes by PCR-SSCP of ITS-2 rDNA

被引:75
作者
Gasser, RB
Monti, JR
机构
[1] Department of Veterinary Science, University of Melbourne, Werribee, Vic. 3030
基金
澳大利亚研究理事会;
关键词
parasitic nematodes (order Strongylida); polymerase chain reaction-linked single strand conformation polymorphism (PCR-SSCP); species identification; second internal transcribed spacer (ITS-2); ribosomal DNA;
D O I
10.1006/mcpr.1997.0106
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Fourteen species of parasitic nematodes (order Strongylida) were characterized using a polymerase chain reaction-linked single strand conformation polymorphism technique (PCR-SSCP). The rDNA region spanning the second internal transcribed spacer (ITS-2) was amplified from parasite DNA by PCR. The PCR products were then denatured and subjected to electrophoresis on a nondenaturing gel matrix. PCR-SSCP of the single stranded ITS-2 molecules generated characteristic and reproducible patterns for each species, and allowed the rapid delineation of all of the 14 species in one step. The method also allowed the display of variation in patterns within some species between different geographical isolates. These findings demonstrate the usefulness of PCR-SSCP of ITS-2 for the rapid identification of nematode species and indicate its potential for resolving variation in the ITS-2 sequence within a species. (C) 1997 Academic Press Limited.
引用
收藏
页码:201 / 209
页数:9
相关论文
共 24 条
[1]   A MOLECULAR PHYLOGENY OF THE GENUS ECHINOCOCCUS [J].
BOWLES, J ;
BLAIR, D ;
MCMANUS, DP .
PARASITOLOGY, 1995, 110 :317-328
[2]   DIFFERENCES IN A RIBOSOMAL DNA-SEQUENCE OF STRONGYLUS SPECIES ALLOWS IDENTIFICATION OF SINGLE EGGS [J].
CAMPBELL, AJD ;
GASSER, RB ;
CHILTON, NB .
INTERNATIONAL JOURNAL FOR PARASITOLOGY, 1995, 25 (03) :359-365
[3]   DIFFERENCES IN A RIBOSOMAL DNA-SEQUENCE OF MORPHOLOGICALLY INDISTINGUISHABLE SPECIES WITHIN THE HYPODONTUS-MACROPI COMPLEX (NEMATODA, STRONGYLOIDEA) [J].
CHILTON, NB ;
GASSER, RB ;
BEVERIDGE, I .
INTERNATIONAL JOURNAL FOR PARASITOLOGY, 1995, 25 (05) :647-651
[4]   CURRENT METHODS OF MUTATION DETECTION [J].
COTTON, RGH .
MUTATION RESEARCH, 1993, 285 (01) :125-144
[5]  
ELLSWORTH DL, 1993, BIOTECHNIQUES, V14, P214
[6]   Fingerprinting sequence variation in ribosomal DNA of parasites by DGGE [J].
Gasser, R ;
Nansen, P ;
Guldberg, P .
MOLECULAR AND CELLULAR PROBES, 1996, 10 (02) :99-105
[7]   Species markers for equine strongyles detected in intergenic rDNA by PCR-RFLP [J].
Gasser, RB ;
Stevenson, LA ;
Chilton, NB ;
Nansen, P ;
Bucknell, DG ;
Beveridge, I .
MOLECULAR AND CELLULAR PROBES, 1996, 10 (05) :371-378
[8]   RAPID SEQUENCING OF RDNA FROM SINGLE WORMS AND EGGS OF PARASITIC HELMINTHS [J].
GASSER, RB ;
CHILTON, NB ;
HOSTE, H ;
BEVERIDGE, I .
NUCLEIC ACIDS RESEARCH, 1993, 21 (10) :2525-2526
[9]   THE EVOLUTION OF EUKARYOTIC RIBOSOMAL DNA [J].
GERBI, SA .
BIOSYSTEMS, 1986, 19 (04) :247-258
[10]  
Hayashi K, 1991, PCR Methods Appl, V1, P34