Sensitive detection of canine parvovirus DNA by the nested polymerase chain reaction

Hirasawa, Tsutomu; Kaneshige, Toshihiko; Mikazuki, Katsumi

doi:10.1016/0378-1135(94)90143-0

Cited by 28 publications

(33 citation statements)

References 31 publications

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“…2). The results were in agreement with those reported by Hirasawa et al [7] in Japan, Steniel et al [19] in South Africa, Pereira et al [14] in Brazil, Wang et al [24] in Taiwan and Hong et al [8] in US. In a recent report Sanjukta et al [17] also reported CPV-2b as the prevalent strain in certain North Indian states.…”

supporting

confidence: 83%

Molecular Typing of Canine parvovirus Occurring in Pondicherry by Multiplex PCR and PCR–RFLP

Parthiban¹,

Mukhopadhyay²,

Antony³

et al. 2010

Indian J. Virol.

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The present study was aimed at molecular typing of Canine parvovirus (CPV) occurring in Pondicherry using PCR based assays. CPV-2a and CPV-2b types were detected by PCR in 68 (53.12%) out of 128 faecal samples/rectal swabs. All the 68 samples found positive by PCR assay were subjected to multiplex PCR assay with CPV-2ab and CPV-2b primer pairs. Sixty-seven (98.52%) samples were characterized as CPV-2b type and one sample (1.47%) was categorized as CPV-2a type. Sixty clinical samples found negative by CPV-2ab primers were subjected to another PCR assay with CPV-555 primer pair for detecting CPV-2c. Though three samples (5%) responded to this PCR, subsequent RFLP of these PCR products with MboII did not show any cleavage indicating the absence of CPV-2c in Pondicherry. It was inferred that CPV-2b was the most prevalent CPV type in Pondicherry. It was further concluded that the CPV-2 variants (CPV-2a, CPV-2b and CPV-2c) currently circulating in the field worldwide could be diagnosed by employing multiplex PCR and PCR-RFLP assays.

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confidence: 83%

Molecular Typing of Canine parvovirus Occurring in Pondicherry by Multiplex PCR and PCR–RFLP

Parthiban¹,

Mukhopadhyay²,

Antony³

et al. 2010

Indian J. Virol.

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“…The number of the genome copy in positive samples was estimated about 10 9 -10 11 /g of faeces by the conventional PCR and 10 11 -10 13 /g of faeces by the nested PCR. Thus, the nested PCR seems to be a sensitive, specific and practical method for the detection of CPV in faecal samples [31,71].…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

“…The PCR can now be used to differentiate the different mutants of CPV-2 using the primers specific for particular mutants [71]. To increase the sensitivity and specificity of the reaction, the nested PCR has been employed [31]. The conventional PCR could detect 10 fg of viral replicative form (RF) DNA on agarose gel electrophoresis, whereas as little as 100 ag of the RF DNA was detected by the nested PCR, which was shown to be 100 times more sensitive than the single PCR [31].…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

“…To increase the sensitivity and specificity of the reaction, the nested PCR has been employed [31]. The conventional PCR could detect 10 fg of viral replicative form (RF) DNA on agarose gel electrophoresis, whereas as little as 100 ag of the RF DNA was detected by the nested PCR, which was shown to be 100 times more sensitive than the single PCR [31]. The number of the genome copy in positive samples was estimated about 10 9 -10 11 /g of faeces by the conventional PCR and 10 11 -10 13 /g of faeces by the nested PCR.…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

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Canine Parvovirus: Current Perspective

2010

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“…M38245). The PCR amplification was performed as previously described with some modifications [12]. The sequences were directly determined from PCR products by the dideoxynucleotide chain termination method with the primer pair A [ Korean isolates belong to CPV-2a were subclassified according to the following categories: (i) all the type 2a isolates had Ser297Ala substitution; (ii) subcluster 2a-I isolates (GenBank accession no.…”

mentioning

confidence: 99%

Genetic Analysis of VP2 Gene of Canine Parvovirus Isolates in Korea

Jeoung

Ahn

Kim

2008

The Journal of Veterinary Medical Science

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ABSTRACT. The purpose of this study was to genetically characterize CPV isolates from Korea. The VP2 gene of 31 isolates was characterized by DNA sequencing and their phylogeny. Among the 31 field CPV isolates, 28 isolates were classified as type 2a and other 3 isolates as type 2b. The isolates in 2a-I, II and III subclusters have unique mutations. The isolates in 2a-IV and V subclusters had similar amino acid sequences to type 2a isolates from other parts of the world. The isolates in type 2b had similar amino acid sequences to type 2b isolates from Asia, Italy, and U.S.A. The molecular analysis of VP2 gene of CPV provided the useful information for the identification of CPV types and the understanding of their genetic relationship.

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Sensitive detection of canine parvovirus DNA by the nested polymerase chain reaction

Cited by 28 publications

References 31 publications

Molecular Typing of Canine parvovirus Occurring in Pondicherry by Multiplex PCR and PCR–RFLP

Molecular Typing of Canine parvovirus Occurring in Pondicherry by Multiplex PCR and PCR–RFLP

Canine Parvovirus: Current Perspective

Genetic Analysis of VP2 Gene of Canine Parvovirus Isolates in Korea

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