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

来源：

INTERNATIONAL JOURNAL FOR PARASITOLOGY | 1999年 / 29卷 / 10期

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

页码：1589 / 1596

页数：8

共 30 条

[21]

MOERKERK PT, 1990, VIRCHOWS ARCH B, V58, P351

[22]

Muller N, 1996, J CLIN MICROBIOL, V34, P2850

[23] Detection of Neospora caninum DNA by the polymerase chain reaction [J].

Payne, S ;

Ellis, J .

INTERNATIONAL JOURNAL FOR PARASITOLOGY, 1996, 26 (04) :347-351

[24] SHEDDING LIGHT ON PCR CONTAMINATION [J].

SARKAR, G ;

SOMMER, SS .

NATURE, 1990, 343 (6253) :27-27

[25]

Shibta Darryl, 1994, P47

[26]

Thilsted J P, 1989, J Vet Diagn Invest, V1, P205

[27] NEOSPORA ABORTION IN NEW-ZEALAND CATTLE [J].

THORNTON, RN ;

THOMPSON, EJ ;

DUBEY, JP .

NEW ZEALAND VETERINARY JOURNAL, 1991, 39 (04) :129-133

[28] A SINGLE TUBE NESTED PCR FOR THE DETECTION OF HEPATITIS-C VIRUS-RNA [J].

TILSTON, P ;

CORBITT, G .

JOURNAL OF VIROLOGICAL METHODS, 1995, 53 (01) :121-129

[29] PREVALENCE OF ANTIBODIES TO NEOSPORA-CANINUM IN A POPULATION OF URBAN DOGS IN ENGLAND [J].

TREES, AJ ;

GUY, F ;

TENNANT, BJ ;

BALFOUR, AH ;

DUBEY, JP .

VETERINARY RECORD, 1993, 132 (06) :125-126

[30] Neospora caninum: Specific oligonucleotide primers for the detection of brain ''cyst'' DNA of experimentally infected nude mice by the polymerase chain reaction (PCR) [J].

Yamage, M ;

Flechtner, O ;

Gottstein, B .

JOURNAL OF PARASITOLOGY, 1996, 82 (02) :272-279

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