Real-time fluorescence RPA reagent kit and test strip RPA reagent kit for rapidly detecting porcine parvovirus and application of reagent kits
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A detection kit and parvovirus technology are applied in the field of rapid detection of porcine parvovirus to achieve the effects of being easy to carry, saving test time and having a simple method.
Active Publication Date: 2016-05-25
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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[0010] For the NS1 gene of PPV, the present invention establishes and evaluates the RPA (real-time RPA) detection kit based on fluorescent probe and the LFSRPA detection kit based on test strips to realize the purpose of rapid detection of PPV. As far as we know, domestic No such kit has been established for the detection of PPV
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Embodiment 1
[0054] Example 1 Establishment of a real-time fluorescent RPA (Real-timeRPA) detection kit and detection method for rapid detection of porcine parvovirus
[0055] 1. Design and preparation of primers and probe sequences
[0056] The inventors of the present invention passed KF429255.1, JX871883.1, JN872448.1, EU790642.1, EU790641.1, L23427.1, M38367.1, M32787.1, D00623.1, KF429254.1 from GenBank , JQ710893.1, KF429252.1, JQ710888.1 and AY502114.1 NS1 gene homologous sequences were compared and analyzed, and the conserved region of porcine parvovirus NS1 gene was determined, and primers and probes were designed for this region, in order to be able to As many porcine parvoviruses as possible were detected. All primers and probes were synthesized by Sangon Biotech (Shanghai, China). The present invention designed and synthesized three pairs of upstream primers and three pairs of downstream primers respectively, as shown in Table 1 below.
[0057] Table 1, PPVReal-timeRPA prime...
Embodiment 2
[0092] Example 2 Establishment of the test strip RPA (LFSRPA) detection kit and detection method for rapid detection of porcine parvovirus
[0093] 1. Design and preparation of primers and probe sequences
[0094]The inventors of the present invention passed KF429255.1, JX871883.1, JN872448.1, EU790642.1, EU790641.1, L23427.1, M38367.1, M32787.1, D00623.1, KF429254.1 from GenBank , JQ710893.1, KF429252.1, JQ710888.1 and AY502114.1 NS1 gene homologous sequences were compared and analyzed, and the conserved region of porcine parvovirus NS1 gene was determined, and primers and probes were designed for this region, in order to be able to As many porcine parvoviruses as possible were detected. All primers and probes were synthesized by Sangon Biotech (Shanghai, China). The present invention designed and synthesized three pairs of upstream primers and three pairs of downstream primers respectively, as shown in Table 4 below.
[0095] Table 4, PPVRPA primers and probes designed by...
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Abstract
The invention discloses a real-time fluorescence RPA (PPV real-time RPA) reagent kit for rapidly detecting porcine parvovirus (PPV) and application of the real-time fluorescence RPA reagent kit, and further discloses a test strip RPA (LFS RPA) reagent kit for rapidly detecting PPV and application of the test strip RPA reagent kit. The real-time fluorescence RPA reagent kit includes a primer shown in SEQ NO.1 and a probe, and the test strip RPA reagent kit includes a primer shown in SEQ NO.2 and a probe, wherein modification basic groups and detection platforms of the primers and the probes are different although the two kinds of reagent kits are oriented to the same target sequence. It is proved through experiments that the sensibility of the two reagent kits is both 102 copy / response, and the two reagent kits can only specifically detect PPV. The two reagent kits are used for detecting a suspected PPV sample, and it shown through the results that the detection results of the two reagent kits have high conformity with qPCR. Thus, the two reagent kits can both rapidly, efficiently and sensitively detect PPV, and an effective technological means is provided for differential diagnosis of PPV.
Description
technical field [0001] The invention relates to a kit for detecting porcine parvovirus and its application, in particular to a real-time fluorescent RPA kit and test strip RPA kit for rapid detection of porcine parvovirus based on RPA reaction, and also relates to the use of both The invention relates to the rapid detection of porcine parvovirus, and the invention belongs to the field of preventive veterinary inspection. Background technique [0002] Thirty years have passed since the birth of PCR technology. From classic PCR, real-time quantitative PCR to the current digital PCR, this technology is constantly changing but has never faded out of our field of vision. However, this technology requires special and expensive thermal cycle instruments and skilled operators, which is time-consuming and labor-intensive, and it is difficult to be used in on-site diagnosis and places with poor experimental conditions. In the laboratories of developing countries, due to the limitati...
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