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Real-time fluorescent quantitative RT-PCR detection primers for serotype of Palyam serogroup virus (PALV), probes and detection kit

A real-time fluorescence quantitative, Paliam technology, applied in the direction of microbial determination/inspection, microorganisms, microorganism-based methods, etc., to achieve the effect of shortening time, saving labor costs, and improving biosafety

Active Publication Date: 2020-01-10
YUNNAN ANIMAL SCI & VETERINARY INST
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Problems solved by technology

[0006] Fluorescent quantitative PCR technology is a new type of nucleic acid qualitative and quantitative technology introduced by ABI Company in the United States in 1996. Since its invention, this technology has been widely used in the detection of bacteria, viruses and other pathogens. It has strong characteristics, high sensitivity and fast detection speed. , high-throughput detection and other advantages, and more importantly, this technology can be used for clinical sample detection, but there is no one-step real-time fluorescent quantitative RT-PCR (one step quantitative real time RT-PCR) that can identify PALV serotypes. , qRT-PCR) detection method available

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  • Real-time fluorescent quantitative RT-PCR detection primers for serotype of Palyam serogroup virus (PALV), probes and detection kit
  • Real-time fluorescent quantitative RT-PCR detection primers for serotype of Palyam serogroup virus (PALV), probes and detection kit
  • Real-time fluorescent quantitative RT-PCR detection primers for serotype of Palyam serogroup virus (PALV), probes and detection kit

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Embodiment Construction

[0061] The present invention will be further described in detail below in conjunction with the examples.

[0062] Those skilled in the art will understand that the following examples are only for illustrating the present invention and should not be considered as limiting the scope of the present invention. If no specific technique or condition is indicated in the examples, it shall be carried out according to the technique or condition described in the literature in this field or according to the product specification. The materials or equipment used are not indicated by the manufacturer, and they are all conventional products that can be obtained through purchase.

[0063] (1) Experimental materials

[0064]PALV CHUV serotype, BCV serotype and DAV serotype strains (19 strains in total) were isolated and sequenced from Paliam serogroup viruses recorded in southern China from 2012 to 2016, Yang Heng, Xiao Lei, Li Zhanhong, Meng Jinxin, Yang Zhenxing, Lu Minna, Lin Xuhui, Liao...

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Abstract

The invention relates to real-time fluorescent quantitative RT-PCR detection primers for a serotype of a Palyam serogroup virus (PALV), probes and a detection kit and belongs to the technical field ofmolecular biological detection on animal viruses. The kit comprises 3 pairs of primers and further comprises three probes, a negative control template, positive control templates, standard substancetemplates and a PCR amplification reagent. RNase-free water serves as the negative control template; the positive control templates are CHUV, BCV and DAV inactivated viruses separately; and the standard substance templates are gene segment 2 single-stranded RNAs of CHUV, BCV and DAV. The kit disclosed by the invention can be used for detecting three kinds of serotype PALVs popular in China and hasthe advantages of specificity, sensitivity, rapidness, high efficiency and the like; and a standard curve established by utilizing the standard substance templates in a matched manner can be used forcarrying out quantitative detection on RNAs of all PALV serotype strains in clinical samples, and thus, the working efficiency is increased greatly.

Description

technical field [0001] The invention belongs to the technical field of animal virus molecular biology detection, and in particular relates to three pairs of amplification primers and three TaqMan probes for rapid detection of the serotype of Paliam serogroup virus popular in China by using TaqMan real-time fluorescent quantitative RT-PCR technology. Needle and assembled test kit. Background technique [0002] Palyam serogroup virus (PALV) is a member of the genus Orbivirus of the Reoviridae family and is widely prevalent in tropical and subtropical regions between 49°N and 35°S. . PALV is mainly transmitted through the blood-sucking bites of female blood-sucking insects Culicoides to animals, infecting cattle and sheep and other ruminants, resulting in abnormal production of pregnant animals, mainly manifested as abortion, premature birth, stillbirth or anencephaly. From 1985 to 1986, the Paliam virus epidemic broke out in Japan. Its clinical symptoms were congenital abnor...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6851C12Q2561/113C12Q2561/101C12Q2521/107Y02A50/30
Inventor 杨恒廖德芳李卓然李占鸿杨振兴肖雷李华春
Owner YUNNAN ANIMAL SCI & VETERINARY INST
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