Development of a single tube nested polymerase chain reaction assay for the detection of Neospora caninum DNA

被引:50
作者
Ellis, JT [1 ]
McMillan, D
Ryce, C
Payne, S
Atkinson, R
Harper, PAW
机构
[1] Univ Technol Sydney, Dept Cell & Mol Biol, Gore Hill, NSW 2065, Australia
[2] Grafton Agr Res & Advisory Stn NSW Agr, Grafton, NSW, Australia
基金
澳大利亚研究理事会;
关键词
Neospora caninum; PCR MIMIC; polymerase chain reaction;
D O I
10.1016/S0020-7519(99)00144-7
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
Sensitive detection techniques are required to study the life cycle of Neospora caninum and to diagnose infections, In this study, we describe the development of a PCR assay for N. caninum based on two successive amplification steps within a single tube. This technique, called single tube nested PCR, was sensitive to a single copy of target sequence, and able to amplify parasite DNA from biological specimens such as formalin-fixed, paraffin-embedded tissues of naturally infected dogs and cattle. An internal standard (or PCR MIMIC) is also described. This assay should prove useful in the study of the biology of N. caninum. (C) 1999 Australian Society for Parasitology Inc. Published by Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:1589 / 1596
页数:8
相关论文
共 30 条
[1]   Comparison of the biological characteristics of two isolates of Neospora caninum [J].
Atkinson, R ;
Harper, PAW ;
Ryce, C ;
Morrison, DA ;
Ellis, JT .
PARASITOLOGY, 1999, 118 :363-370
[2]   ISOLATION OF NEOSPORA-CANINUM FROM A BRITISH DOG [J].
BARBER, J ;
TREES, AJ ;
OWEN, M .
VETERINARY RECORD, 1993, 133 (21) :531-532
[3]  
BJORKMAN C, 1994, ACTA VET SCAND, V35, P445
[4]   BOVINE NEOSPORA ABORTION IN NORTH-EASTERN NEW-SOUTH-WALES [J].
BOULTON, JG ;
GILL, PA ;
COOK, RW ;
FRASER, GC ;
HARPER, P ;
DUBEY, JP .
AUSTRALIAN VETERINARY JOURNAL, 1995, 72 (03) :119-120
[5]   POST-PCR STERILIZATION - A METHOD TO CONTROL CARRYOVER CONTAMINATION FOR THE POLYMERASE CHAIN-REACTION [J].
CIMINO, GD ;
METCHETTE, KC ;
TESSMAN, JW ;
HEARST, JE ;
ISAACS, ST .
NUCLEIC ACIDS RESEARCH, 1991, 19 (01) :99-107
[6]  
DRAGON EA, 1994, DIAGNOSTIC MOL BIOL, P161
[7]  
DUBEY JP, 1988, J AM VET MED ASSOC, V192, P1269
[8]   Comparison of the large subunit ribosomal DNA of Neospora and Toxoplasma and development of a new genetic marker for their differentiation based on the D2 domain [J].
Ellis, JT ;
Amoyal, G ;
Ryce, C ;
Harper, PAW ;
Clough, KA ;
Homan, WL ;
Brindley, PJ .
MOLECULAR AND CELLULAR PROBES, 1998, 12 (01) :1-13
[9]   Polymerase chain reaction approaches for the detection of Neospora caninum and Toxoplasma gondii [J].
Ellis, JT .
INTERNATIONAL JOURNAL FOR PARASITOLOGY, 1998, 28 (07) :1053-1060
[10]   RECENT ADVANCES IN THE POLYMERASE CHAIN-REACTION [J].
ERLICH, HA ;
GELFAND, D ;
SNINSKY, JJ .
SCIENCE, 1991, 252 (5013) :1643-1651