A primer, probe and kit for specific detection of type 2 ungulate boca parvovirus

Abstract

The invention discloses a primer, a probe and a kit for specific detection of type 2 ungulate Boca parvovirus. The primer includes upstream and downstream primers, the nucleotide sequence of the upstream primer PboVF1 is described as SEQ ID NO. 1, the nucleotide sequence of the downstream primer PBoVR1 is SEQ ID NO. 2; the sequence of the probe PboVP1 is shown as SEQ ID NO. 3. The detection kit and reagents have the advantages of accurate detection, high sensitivity, strong specificity, simpleness and rapidness, and have good detection ability.

Description

technical field [0001] The invention relates to biological detection technology, in particular to a primer, a probe and a kit for specifically detecting type 2 ungulate boca parvovirus. Background technique [0002] Bocavirus belongs to the Bocavirus genus of the Parvoviridae subfamily of the Parvoviridae family. Parvoviruses include Thickoviridae and Parvoviridae, the former mainly infecting insects and the latter mainly infecting vertebrates. The Parvovirus subfamily is further divided into 6 genera including Parvovirus, Bocavirus, Rhodovirus, Relivirus, and Mink Aleutian virus. Porcine Bocavirus (Porcine Bocavirus, PBoV, also known as porcine Boca parvovirus) belongs to the Bocavirus genus. It is a single-strand linear non-enveloped DNA virus with a genome of about 5.2kb, including 3 open reading Frames (ORFs), NS1, NP1 and VP1 / VP2, respectively. It mainly attacks the respiratory system and digestive system of pigs. In China, the positive rate of PBoV in pigs with mult...

AI Technical Summary

Problems solved by technology

However, since the VP1 / VP2 genes are widely present in porcine bocaviruses, the above method cannot diagnose the three genotypes under the type 2 ungulate bocaparvovirus (Ungulate bocaparvovirus 2) population from other porcine bocaviruses , and different populations of porcine bocaviruses may have differences in the infected tissue, pathogenic mechanism and host selection, and the diagnostic accuracy is low and it is easy to delay the timing of treatment

In addition, due to the high mutation rate of the porcine bocavirus gene sequence, it is difficult to design a fluorescent quantitative PCR detection method for boca parvovirus in ungulates type 2 through conventional techniques

The complete VP1 / VP2 gene has insufficient conservation and stability in these three genotypes, and many sites are prone to mutations

Qin Shaomin et al. (Qin Shaomin, Wu Jianmin, Genotyping Research 150-153_ Ma Lin, et al. Sequence Analysis and Genotyping Research of Porcine Boca Virus Whole Genome[J]. Chinese Journal of Preventive Veterinary Medicine, 2014 36(2)) passed the research 28 complete gene sequences of porcine Boca virus, found that gene insertion is common in porcine Boca virus, and the mutation rate is very high, the homology of the whole gene sequence is between 9.3% and 97.5%, and there is genetic diversity

Especially in the conventional in vitro PCR amplification process, if the copy number of the VP1 / VP2 gene in the sample is small and multiple cycles are required for amplification, multiple sites of the VP1 / VP2 gene in the amplified product are affected by the reaction system. The impact will change, and the coefficient of variation is as high as 5.44%-8.61% after testing, resulting in the diversity of the amplified target genes, and the fluorescent probes for VP1 / VP2 genes cannot bind to the amplified target fragments

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