RT-PCR primers for detecting mink astroviruses and detection method using the same

A RT-PCR and astrovirus technology, which is applied in the field of RT-PCR primers and its detection for the detection of mink astrovirus, can solve the problems of lack of literature related to mink astrovirus, and achieve high sensitivity, low cost and suitable length Effect

Active Publication Date: 2016-01-20
QINGDAO AGRI UNIV
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Problems solved by technology

However, at present in our country, there are still relatively few studies on mink astrovirus, and the relevant literature on mink astrovirus is relatively scarce.

Method used

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  • RT-PCR primers for detecting mink astroviruses and detection method using the same
  • RT-PCR primers for detecting mink astroviruses and detection method using the same
  • RT-PCR primers for detecting mink astroviruses and detection method using the same

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Embodiment

[0029] The mink astrovirus-positive disease materials used in the examples of the present invention; mink parvovirus; mink distemper virus; porcine epidemic diarrhea virus are all preserved by the Key Laboratory of Preventive Veterinary Medicine of Qingdao Agricultural University.

[0030] Step 1, design and synthesis of primers: compare the nucleotide sequence ( figure 1 ), the relatively conserved region of the selected gene. Utilize Primeer5.0 software, take mink astrovirus (NC_004579.1) ORF1b gene sequence as a reference, design the specific primer that detects mink astrovirus ( figure 2 ,Table 1). The primers were synthesized by Beijing Saibaisheng Gene Technology Co., Ltd., diluted with DEPC water to 10 pmol / ul, and stored at -20 degrees Celsius for later use.

[0031] Table 1 Primer Sequence

[0032]

[0033] Note: Position in the whole genome of Minkastrovirus strain (NC_004579.1)

[0034] Step 2: Use TaKaRa's RNAisoPlus reagent to extract total RNA in the samp...

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Abstract

The invention discloses RT-PCR primers for detecting mink astroviruses and a detection method using the same. According to a mink astrovirus ORF1b gene sequence (with serial numbers of GU985458.1, NC_004579.1 and AY179509.1) conservation region in the GenBank, a pair of specific primers are analyzed and designed, according to the designed primers, the optimal reaction system and reaction conditions are researched, a RT-PCR detection method is built and a specificity test, a sensitivity test and a repeatability test are carried out. Test results shows that the mink astrovirus RT-PCR method has strong sensitivity and high specificity and can realize fast detection of mink astroviruses. The mink astrovirus RT-PCR method has good applicability and can be used for detecting mink astroviruses.

Description

technical field [0001] The invention relates to the field of molecular biology molecular markers, in particular to an RT-PCR primer for detecting mink astrovirus and a detection method thereof. Background technique [0002] Mink astrovirus (MiAstV) is one of the pathogenic causes of mink diarrhea symptoms, especially in mink around weaning and juvenile mink, the incidence of astrovirus is more prominent. However, at present in our country, there are still relatively few studies on mink astrovirus, and the relevant literature on mink astrovirus is relatively scarce. From 2011 to 2013, in some mink breeding areas in my country, a central nervous system disorder disease appeared in young minks aged 1 to 2 months. It was found through research that the disease was related to mink tremor syndrome in Sweden, Denmark and other countries. Symptoms of Shankingmink syndrome (SMS) are similar. The main symptoms are head tremor, gait instability, and ataxia. Later, it develops into hind...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/701
Inventor 黄娟单虎袁飞
Owner QINGDAO AGRI UNIV
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