Nucleic acid reagents, kits and systems for detecting enteroviruses
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An enterovirus and kit technology, which is applied in the field of enterovirus detection, can solve the problems of complex sample processing, increase operational complexity, and judge complex results, achieve simple detection process, avoid sample extraction steps, and automatic results. interpret and effect
Active Publication Date: 2019-10-08
北京卓诚惠生生物科技股份有限公司
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Commonly used PCR detection requires complex sample processing, cumbersome test procedures and complex result judgments
In addition, the detection targets that RT-PCR can cover are limited, and multi-system settings are required to achieve multi-target detection, which increases the complexity of the operation
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[0048] 1. Primer and probe synthesis
[0049] Sequence synthesis was performed according to the primer and probe sequences shown in Table 1 and Table 2. In the sequence, Y represents the degenerate base T / C; R represents the degenerate base A / G; W represents the degenerate base A / T; FAM in the probe is 6-carboxyfluorescein, JOE is 2,7-di Methyl-4,5-dichloro-6-carboxyfluorescein, TAMRA is 6-carboxytetramethylrhodamine, CY5 is 5H-indocyanine. The brackets in the probe sequences in Table 2 indicate that the t on the left side of the brackets has a fluorescent label, and the contents in the brackets indicate the choice of fluorescent labels.
[0050] Table 1
[0051]
[0052]
[0053] Table 2
[0054]
[0055] 2. Template extraction
[0056]After collecting clinical samples such as nasopharyngeal swabs with the sampler matched with ParaDNA, they can be amplified directly in the ParaDNA reactor.
[0057] 3. Construct Hybeacon probe technology detection system
[0058...
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Abstract
The present disclosure relates to a nucleic acid reagent, kit and system for detecting enteroviruses. The nucleic acid reagent comprises primers shown in SEQ ID NO.1-36 and probes shown in SEQ ID NO.39-58, wherein the primers and probes are stored separately from each other or arbitrarily mixed with each other. The nucleic acid reagent, kit and system detecting 22 pathogenic enteroviruses are established by using the above primers and probes, and can realize rapid, comprehensive, sensitive, specific and automatic detection result determination, and significantly improve sensitivity, specificity, and simplicity of simultaneous detection of multiple enteroviruses.
Description
technical field [0001] The present disclosure relates to the detection of enteroviruses, in particular to a nucleic acid reagent, kit and system for detecting enteroviruses. Background technique [0002] Enteroviruses belong to the genus Enterovirus in the family Picornaviridae. There are many kinds of enteroviruses that can cause human pathogenicity, including human enterovirus A, B, C, and D groups, with more than 100 serotypes. Common human enteroviruses mainly include: Coxsackievirus A (Coxsackievirus A, CA, common types include types 2, 4, 5, 6, 7, 9, 10, 12, 16), Coxsackievirus B Group (Coxsackievirus B, CB, common types include types 1, 2, 3, 4, 5), human intestinal cytopathic orphan virus (Echovirus ECHO), new enterovirus, etc. New enteroviruses were isolated successively after 1969, such as novel enterovirus 71 (EV71). At present, domestic studies on subtypes of CA16 and EV71 are also increasing day by day, and it is of great significance to understand the preval...
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