Norovirus nucleic acid detection kit and use method thereof

A technology for detecting kits and viral nucleic acids, which is applied in the field of norovirus nucleic acid detection, can solve the problems of easy error cost and slow efficiency, and achieve the effects of improving stability, ease of use, and improving sensitivity

Active Publication Date: 2022-01-14
SHANGHAI BIOGERM MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention provides a norovirus nucleic acid detection kit and its use method, aiming at the technical problems of slow detection of norovirus GⅠ and GⅡ virus detection and typing and identification, easy error and high cost

Method used

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  • Norovirus nucleic acid detection kit and use method thereof
  • Norovirus nucleic acid detection kit and use method thereof
  • Norovirus nucleic acid detection kit and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0113] Norgin virus's classification effect verification: If a Nor is a virus GI (FAM fluorescent group mark, BHQ1 quenching group mark) and GII type (Vic fluorescent group mark, BHQ1 quenching group mark) probe setting Different passages, the kit can be developed as a non-virus GI and GII type typing test reagents, for 10 samples (inactivated clinical samples from GI infection from Shanghai CDC), No. 10) and No. 11 sample (From the inactivation clinical sample of GII infection from Shanghai CDC, No. 11) is detected, and the GI-type and GII-type amplification curves correspond to different fluorescence channels, indicating that fluorescence of corresponding and label probes. Channel, can effectively distinguish between GI and GII types, as shown figure 1 .

Embodiment 3

[0115] Detection and precision verification of clinical samples: The Norgin Virus GI is set to the GII probe, and the kit is set to a non-Virus GI and GII-type universal detection reagents, for 74 cases of clinical samples (originating from The GI and / or GII-type inventive clinical samples of the Shanghai CDC, numbered 1-74, Table 2), and diluted to 50 copies / ml (detection limit concentration) to be detected. The detection results of Table 2 can be seen that the kit of the present invention can effectively detect the non-virus GI and GII type samples of the detection limit concentration, and the detection rate is 100%, indicating that it is 50 copies / ml for clinical samples. The results of its detection are shown in Figure 2. Dilute 10-20 samples to 100 copies / ml (low concentration) and 10 5 COPIES / ML (high concentration), its amplification curve has no abnormalities, CT value of CV is 0.8% and 0.6%, and it is less than 5%, indicating that its precision is qualified, and...

Embodiment 4

[0119] Specificity Verification (Cross Reaction): The specific detection effect of other pathogens is detected in Table 3. It is negative that other common pathogens are detected.

[0120]

[0121]

[0122] table 3

[0123] Specific verification (anti-interference): The kit detection control group and the addition of different interference substances were detected, and the comparative test results of the Nori virus Gi type were added, and the comparative test results of the interference substance and the control group (non-interference substance) were shown in Table 4. . The results show that when the Norgin Virus GI sample contains the interference material in the following table, there is no interference to the kit, indicating that the reagent box anti-interference ability is strong.

[0124]

[0125]

[0126] Table 4: Novoli virus GI interference substance research experimental data

[0127] Specific verification (anti-interference): The kit is detected the control grou...

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Abstract

The invention discloses a norovirus universal nucleic acid detection kit and a use method thereof. The norovirus universal nucleic acid detection kit comprises a norovirus reaction solution, a positive control solution and a negative control solution, and the norovirus reaction solution is freeze-dried powder which is pre-packaged into eight connecting tubes, wherein the norovirus reaction solution contains a plurality of probes with different fluorescence labels, primers, an enzyme mixture, an enhancer, a freeze-drying protective additive, a 10 * buffer, a nucleotide mixture and the like, and whether the norovirus is infected or not can be detected in the same reaction system. The whole process of sample collection, nucleic acid extraction, amplification and detection is monitored through the endogenous internal reference, and misjudgment caused by false negative results is avoided. The probes are oligonucleotide comprising a fluorescence reporter group and a quenching group. When the probes are complete, the quenching group is close to the reporter group in spatial position, so that fluorescence emitted by the reporter group is inhibited. When the primers extend, the probe combined with the template is cut off by Taq enzyme (5 '-> 3' exonuclease activity), the reporter group is separated from the quenching group, and a fluorescence signal is generated, so that the detection of the norovirus on the nucleic acid level is realized.

Description

Technical field [0001] The present invention relates to the field of nor, viral nucleic acid detection, particularly in a non-viral nucleic acid detection kit and method thereof. Background technique [0002] Norovirus (NV) is an RNA virus that causes intestinal acute disorders, according to the similarity of nucleotide sequences in the RNA polysesence zone, and divides NV into 5 genotypes (GI ~ GV), including GI, GII and GIV can be infected. Norgin virus has been considered by many countries to lead to the first pathogens of adult and child viral diarrhea and gastroenteritis, which are mainly in-manure-mouth, followed by direct contact infection, and China's popular human Nuo, like viruses mainly in Genome II as host. The clinical manifestation of infection is acute gastroenteritis and permeability of diarrhea. The course of disease is generally 7 days, and the fever lasts for 3 days, vomiting 2 ~ 3 days, diarrhea 5 days, severe dehydration symptoms. Nor is like a virus in cold ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/166C12Q2563/107Y02A50/30
Inventor 赵百慧杨孙孝李春明王馨玉
Owner SHANGHAI BIOGERM MEDICAL TECH CO LTD
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