Detection of porcine parvovirus DNA by the polymerase chain reaction assay using primers to the highly conserved nonstructural protein gene, NS-1.
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.
Soares
R M
RM
Laboratory of Applied Molecular Biology, Faculty of Veterinary Medicine and Zootechny, University of São Paulo, Brazil.
Durigon
E L
EL
Bersano
J G
JG
Richtzenhain
L J
LJ
D000034:Abortion, Veterinary / Q000821:virology; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D005313:Fetal Death / Q000662:veterinary; D007668:Kidney / Q000166:cytology; D008969:Molecular Sequence Data; D010322:Parvoviridae Infections / Q000662:veterinary* / Q000821:virology; D017992:Parvovirus / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D013552:Swine; D013553:Swine Diseases / Q000821:virology*; D017361:Viral Nonstructural Proteins / Q000235:genetics*
D017931:DNA Primers; D004279:DNA, Viral; C106202:NS1 protein, parvovirus; D017361:Viral Nonstructural Proteins