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LAMP detection primer group and kit for white spot syndrome virus (WSSV)

A technology of leukoplakia syndrome and a detection kit, which is applied in the field of molecular biology, can solve problems affecting the judgment of results, and achieve the effects of simple operation, high specificity, and easy identification

Active Publication Date: 2015-03-04
广州迪澳生物科技有限公司 +1
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0005]Because the ESE-Quant tube scanner judges whether the reaction is progressing by absorbing fluorescence, the previous research found that the commonly used fluorescent color reagent SYBR Green I is too sensitive, and in ESE-Quant Non-specific amplification curves often appear in negative reaction wells on the tube scanner, which affects the judgment of results

Method used

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  • LAMP detection primer group and kit for white spot syndrome virus (WSSV)
  • LAMP detection primer group and kit for white spot syndrome virus (WSSV)
  • LAMP detection primer group and kit for white spot syndrome virus (WSSV)

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Embodiment 1

[0046] 1. Establishment of LAMP detection kit for shrimp white spot syndrome virus

[0047] LAMP detection kit for shrimp white spot syndrome virus, including LAMP primer set, LAMP reaction solution, Bst DNA polymerase, positive and negative controls.

[0048] 1) LAMP primer design: According to the nucleic acid sequence of shrimp white spot syndrome virus (WSSV) retrieved from Genbank, select a conserved sequence and design specific primers for detecting WSSV. The sequence is shown in Table 1 below:

[0049] Table 1 Primer sequence list

[0050]

[0051] 2) LAMP reaction solution: mix 10mM dNTP, 10×ThermoPol reaction buffer, 150mM MgSO 4 The aqueous solution and the three are mixed in a volume ratio of 8:5:2.

[0052] 3) The positive control is a T vector containing the target sequence fragment, and its preparation method is: using the isolated and identified shrimp white spot syndrome virus DNA as a template, using the external primers in Table 1 (SEQ ID NO: 1 and SE...

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Abstract

The invention discloses a WSSV prawn white spot syndrome virus (WSSV) LAMP detection primer set and a detection kit. The detection primer set includes a pair of outer primers, a pair of inner primers and a pair of loop primers; the detection kit includes a primer solution, a reaction solution, DNA polymerase and a reference substance; the kit also includes a chromogenic reagent. The kit of the invention is easy and quick to operate, suitable for on-site detection, strong in specificity, high in detection sensitivity up to 1 fg / μL, high in accuracy and reliability, and has wide application prospects.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and relates to a detection method for marine aquaculture, in particular to a LAMP detection primer set and a detection kit for shrimp white spot syndrome virus (WSSV). Background technique [0002] White spot syndrome virus (white spot syndrome virus, WSSV) is the main pathogen of the worldwide shrimp outbreak in 1993, so far there is still no effective technical measures to control the outbreak of WSSV. Scholars at home and abroad generally believe that comprehensive prevention is a relatively effective method, that is, to detect diseases as early as possible and take many measures to prevent the spread of the virus. Therefore, establishing a sensitive, accurate, fast and convenient WSSV pathogen detection method is an important way to reduce the occurrence and harm of white spot syndrome. At present, the detection methods of WSSV pathogens mainly include visual observation, histology,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6844C12Q1/70C12Q2531/119
Inventor 李红梅赵哲邓艳柏建山刘助红唐大运叶蕾张璜石磊
Owner 广州迪澳生物科技有限公司
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