Detection of porcine parvovirus DNA by the polymerase chain reaction assay using primers to the highly conserved nonstructural protein gene, NS-1

被引:67
作者
Soares, RM
Durigon, EL
Bersano, JG
Richtzenhain, LJ [1 ]
机构
[1] Univ Sao Paulo, Fac Vet Med & Zootechny, Lab Appl Mol Biol, Sao Paulo, Brazil
[2] Univ Sao Paulo, Inst Biomed Sci, Virol Lab, Sao Paulo, Brazil
[3] Biol Inst Sao Paulo, Sao Paulo, Brazil
关键词
porcine parvovirus; polymerase chain reaction; detection of viral DNA;
D O I
10.1016/S0166-0934(98)00177-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Porcine parvovirus (PPV) infection is associated with reproductive losses in swine and its causative agent, the PPV, has been isolated worldwide. Serological surveys and virus isolation studies throughout Brazil confirm the occurrence of PPV infection in this country. The most common methods to detect PPV infection are fluorescent antibody staining of fetal tissues, hemagglutination assay of tissue extracts and virus isolation from fetal tissues. Non-specificity and low sensitivity are the major drawbacks of these techniques. The development of a polymerase chain reaction (PCR) and nested-PCR assays for PPV DNA detection from infected cell lines and clinical samples is described. The primers were designed to a highly conserved region of the PPV genome which codes for the non-structural protein, NS-1. Results showed that PCR could detect PPV in titres at least 10(6) higher than the hemagglutination assay. The PCR and nested-PCR assays were used to detect successfully PPV DNA in clinical samples. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:191 / 198
页数:8
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