Canine parvovirus detection kit

Abstract

The invention relates to a canine parvovirus detection kit. The kit comprises 20 PCR reaction tubes, a 300[mu]l PCR reaction mixture, 2.0g of agarose, 100[mu]l of ethidium bromide 10[mu]g / [mu]l, 100[mu]l of 0.25% bromophenol blue sampling buffer solution and 5[mu]l of Taq enzyme (5U / [mu]l), wherein the PCR reaction mixture contains a primer sequence Primer1: F5'-CCAACCATACCAACTCCAT-3', R5'-CTTGTGTAGACGCCTCAA-3', or a Primer 2: F5'-GGAGCAGTTCAACCAGAC-3', R5'-GCATCAACCAATGACCAAG-3'. The canine parvovirus detection kit has the beneficial effects that the primer sequence is highly conservative, canine parvovirus with different sources and mutations can be detected, and the canine parvovirus mutations can be better coped.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a detection kit for canine parvovirus. Background technique [0002] Canine parvovirus disease is an acute and severe infectious disease caused by canine parvovirus (CPV), clinically characterized by vomiting, non-suppurative myocarditis or hemorrhagic enteritis. In 1978, Appel et al. isolated the virus from dogs suffering from enteritis for the first time. The disease was acute and had a high mortality rate. With the rapid growth of the global economy and the continuous improvement of living standards, the number of pets kept increased dramatically. CPV has caused great losses to the kennel industry, seriously endangered the healthy development of the kennel industry, and attracted the attention of many experts at home and abroad. [0003] CPV is a member of the genus Parvovirus in the family Parvoviridae, including feline panleukopenia virus (FPV), raccoon parvovirus (RPV), mink e...

AI Technical Summary

Problems solved by technology

According to the epidemiological characteristics, typical clinical symptoms and pathological changes of canine parvovirus, a preliminary diagnosis can be made. The characteristic symptoms of clinical diagnosis of parvovirus are vomiting, loose stools, and blood in the stool. Puppies often have hemorrhagic enteritis and diarrhea, accompanied by white blood cells. The clinical manifestations are very complicated, but it is difficult to distinguish them from canine distemper, canine coronavirus disease and canine acute gastroenteritis clinically, so it is difficult to use them as the main basis for diagnosis; virus isolation and identification is the most important way to identify viruses The basic method, the primary cells used for culturing CPV include bovine fetal spleen cells, canine small intestine, kidney, spleen and thymocytes, raccoon salivary gland cells, cat fetal kidney cells, cat kidney cells, dog and cat kidney cell lines, mink lung Passage cell lines such as cell lines, because the cell separation cycle is long, at least 1 week, so it is rarely used for clinical detection of CPV; dogs and cats infected with CPV have high levels of virus in feces, serum, and enterovirus, and feces are generally used Electron microscopy is the best way to detect disease, but the experiment is complicated and expensive; CPV HA-HI test is currently the most commonly used method for detecting CPV. The principle is that CPV particles can agglutinate with monkey or pig red blood cells visible to the naked eye. The reaction has the advantages of simple operation, sensitive test, and reliable results. The disadvantage is that porcine red blood cells need to be kept on hand. The disadvantage is that the antibodies produced by the animal body after infection with CPV are quickly combined with antigens, which affects the combination of known antigens and antibodies, resulting in False negative test results; ELISA is a detection method developed after immunofluorescence technology, which can be used to detect antigens or antibodies, including indirect ELISA, antigen competition ELISA, sandwich ELISA and Dot-ELISA, but the sensitivity of ELISA is poor; immunocolloidal gold The diagnostic technology has been widely used in practice because of its gradual and rapid characteristics. This technology is suitable for rapid detection. The main problem at present is that the accuracy still needs to be improved; the sensitivity of FQ-PCR is very high, but the CPV gene often For mutations, positive cases detected by the FQ-PCR method still need to be confirmed by virus isolation and identification, gene sequence comparison analysis and other methods; PCR has become a commonly used diagnostic method because of its high sensitivity, simplicity and speed, and low requirements for the purity of samples. one of the methods

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