A competitive PCR assay for quantitative detection of Neospora caninum

被引：62

作者：

Liddell, S ^{[1
]}

Jenkins, MC ^{[1
]}

Dubey, JP ^{[1
]}

机构：

[1] USDA ARS, Livestock & Poultry Sci Inst, Parasite Biol & Epidemiol Lab, Beltsville, MD 20705 USA

来源：

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

关键词：

competitive quantitative polymerase chain reaction; Neospora caninum;

D O I：

10.1016/S0020-7519(99)00101-0

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：

A quantitative-competitive PCR (QC-PCR) assay was developed for measurement of Neospora caninum levels in the tissues of infected animals. A molecule was synthesised for use in PCR as a competitor to the target Neospora-specific Nc5 genomic sequence. The assay was used to evaluate the relative level of parasites in the brain and lungs of mouse pups in a model of vertical transmission of N. caninum. Infection on day 11 of gestation resulted in similar levels of parasites in all offspring. The assay should be useful in evaluation of vaccines against Neospora infection. Incorporation of the competitor molecule in the detection assay also provides a control for PCR failure and facilitates identification of truly negative samples. (C) 1999 Australian Society for Parasitology Inc. Published by Elsevier Science Ltd. All rights reserved.

引用

页码：1583 / 1587

页数：5

共 24 条

[1]

Anderson ML, 1997, J AM VET MED ASSOC, V210, P1169

[2] Naturally occurring vertical transmission of Neospora caninum in dogs [J].

Barber, JS ;

Trees, AJ .

INTERNATIONAL JOURNAL FOR PARASITOLOGY, 1998, 28 (01) :57-64

[3]

BARR BC, 1994, LAB INVEST, V71, P236

[4] Experimental infection of non-pregnant and pregnant sheep with Neospora caninum [J].

Buxton, D ;

Maley, SW ;

Thomson, KM ;

Trees, AJ ;

Innes, EA .

JOURNAL OF COMPARATIVE PATHOLOGY, 1997, 117 (01) :1-16

[5] The pathogenesis of experimental neosporosis in pregnant sheep [J].

Buxton, D ;

Maley, SW ;

Wright, S ;

Thomson, KM ;

Rae, AG ;

Innes, EA .

JOURNAL OF COMPARATIVE PATHOLOGY, 1998, 118 (04) :267-279

[6] VERTICAL TRANSMISSION OF NEOSPORA-CANINUM IN MICE [J].

COLE, RA ;

LINDSAY, DS ;

BLAGBURN, BL ;

DUBEY, JP .

JOURNAL OF PARASITOLOGY, 1995, 81 (05) :730-732

[7] A review of Neospora caninum and neosporosis [J].

Dubey, JP ;

Lindsay, DS .

VETERINARY PARASITOLOGY, 1996, 67 (1-2) :1-59

[8] Canine neosporosis: clinical signs, diagnosis, treatment and isolation of Neospora caninum in mice and cell culture [J].

Dubey, JP ;

Dorough, KR ;

Jenkins, MC ;

Liddell, S ;

Speer, CA ;

Kwok, OCH ;

Shen, SK .

INTERNATIONAL JOURNAL FOR PARASITOLOGY, 1998, 28 (08) :1293-1304

[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] Detection of Neospora sp. from infected bovine tissues by PCR and probe hybridization [J].

Ho, MSY ;

Barr, BC ;

Rowe, JD ;

Anderson, ML ;

Sverlow, KW ;

Packham, A ;

Marsh, AE ;

Conrad, PA .

JOURNAL OF PARASITOLOGY, 1997, 83 (03) :508-514

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