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Kit for detecting HBV through bulk sample PCR, and application thereof

A technology of kits and reagents, applied in the field of kits for detection of HBV by sample collection PCR, can solve the problems of repeatability and stability, and the cost of kits does not have advantages, so as to ensure the detection rate, reduce sample processing time and Steps, the effect of saving reagent expenses

Inactive Publication Date: 2016-01-27
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, due to the different batches and manufacturers of the kit itself, its repeatability and stability are also affected, and commercialized kits do not have an advantage in terms of cost

Method used

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  • Kit for detecting HBV through bulk sample PCR, and application thereof
  • Kit for detecting HBV through bulk sample PCR, and application thereof
  • Kit for detecting HBV through bulk sample PCR, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1, the design of primer

[0029] According to the comparison of HBV conserved sequences, two pairs of degenerate primers S1-F / S1-R and S2-F / S2-R were designed to establish a nested PCR method for HBV detection. The specific sequences are listed in Table 1;

[0030] Table 1 is the HBVPCR amplification primer sequence

[0031]

Embodiment 2

[0032] Embodiment 2, primers are used for PCR detection of HBV by collection sample method

[0033] 1. Preparation of sample to be tested by collection method

[0034] A total of 32 liver samples from slaughtered chickens were collected, marked and stored at -80°C.

[0035] Put about 100 mg of each of the 4 samples to be tested in a mortar, add liquid nitrogen to grind and mix them evenly, and then use a 1.5ml centrifuge tube to collect samples and mix and grind about 100 mg of tissue for follow-up tests. There are 8 groups of mixed samples to be tested. The mixed samples of each sample set shall be marked for secondary detection.

[0036] One group: numbered 13-16, two groups: numbered 17-20, three groups: numbered 1-4, four groups: numbered 21-24, five groups: numbered 25-28, six groups: numbered 5-8, seven groups: numbered 9-12, eight groups: numbered 29-32. Among the above 32 samples, the samples coded as 1, 5, 8, and 13 were known to be infected with HBV, and the rest ...

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Abstract

The invention discloses a kit for detecting HBV through bulk sample PCR, and an application thereof. The kit is used for identification or assisted identification of hepatitis B virus infected samples in a plurality of samples to be measured, and comprises an individually used primer pair A, an individually used primer pair B and a vector for recording a PCR amplification method. Experiments in the invention prove that PCR detection of collected samples is carried out in HBV epidemiology investigation through a bulk sample method (a sample co-treatment method), so the kit reduces the sample treatment time and step and saves the reagent expenditure especially when the positive rate is not high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting HBV by sample-collecting PCR and its application. Background technique [0002] Hepatitis B (HB) is one of the infectious diseases caused by hepatitis B virus (HBV), which is characterized by liver damage and seriously endangers the health of humans and animals. HBV is a member of the Hepadnaviridae family. So far, the hepadnaviridae discovered have positive hepadnaviruses infecting mammals, which can infect humans and orangutans and other mammals, and are found in animals such as chimpanzees, gibbons, ground squirrels and tree shrews. All have been found; and another type of poultry and birds infected with hepatovirus are also found in Peking duck, snow geese and herons. It is worth noting that in recent years, studies have reported that HBV-like species have also been found in parrots, bats, and slaughtered pigs and chickens. The discovery of HBV in new animal...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 佘锐萍赵月肖鹏毛晶晶杨依霏石蕊寒陈建
Owner CHINA AGRI UNIV
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