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
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
收藏
页码:1583 / 1587
页数:5
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