[Microassay of High-Risk Human Papillomavirus Genotype Based on R6G-ddATP/SNaPshot-Gel Fluorescence Method].
Gel fluorescence method
HR-HPV
R6G-ddATP
SNaPShot technology
Journal
Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition
ISSN: 1672-173X
Titre abrégé: Sichuan Da Xue Xue Bao Yi Xue Ban
Pays: China
ID NLM: 101162609
Informations de publication
Date de publication:
Jan 2021
Jan 2021
Historique:
entrez:
21
1
2021
pubmed:
22
1
2021
medline:
23
1
2021
Statut:
ppublish
Résumé
R6G-ddATP was used as a dideoxy fluorescence substrate to establish the single base end extension (SNaPShot)-gel fluorescence method for the rapid detection of the genotypes of three high-risk human papillomaviruses (HR-HPV) ( HPV quality control products were used as as samples, and R6G-ddATP dideoxy fluorescence reagent was used as substrate. Firstly, HPV was amplified by using universal primers to obtain the first round of amplified products, which were purified and used as templates for subsequent SNaPShot reactions. Then, specific one-step extension primers were used to perform SNaPShot reaction to generate R6G-fluorescence-labeled DNA extension products. The product was subjected to agarose gel electrophoresis, the results of which were observed under a Gel Imager, and the HPV genotyping was done with different one-step extension primers. Each sample was tested three times and the results were compared with DNA sequencing results. The preferred annealing temperature for SNaPShot reaction is 55 ℃. Three The R6G-ddATP/SNaPShot-gel fluorescence method for the micro-detection methods of three HR-HPV genotypes was successfully established and can be used for rapid detection of
Identifiants
pubmed: 33474897
doi: 10.12182/20210160107
pmc: PMC10408940
doi:
Substances chimiques
DNA, Viral
0
Deoxyadenine Nucleotides
0
Dideoxynucleotides
0
2',3'-dideoxyadenosine triphosphate
24027-80-3
Types de publication
Journal Article
Langues
chi
Sous-ensembles de citation
IM
Pagination
98-103Informations de copyright
Copyright© by Editorial Board of Journal of Sichuan University (Medical Sciences).
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