CNV Encompassing SNP Detection by Cross-reactivity Probe
10.13865/j.cnki.cjbmb.2025.04.1100
- VernacularTitle:交叉活性探针确定单核苷酸多态性相关的拷贝数目变异
- Author:
Xiao-Mei HU
1
;
Chen ZHOU
;
Pin-Zheng ZHANG
;
Yang CHEN
;
Jia-Wen LI
;
Yu-Kai MA
;
Jia-Qi WANG
;
Zhi-Yi GUO
Author Information
1. 华北理工大学基础医学院病原生物学系,河北唐山 063200;河北省慢性疾病基础医学重点实验室,河北唐山 063200
- Publication Type:Journal Article
- Keywords:
digital polymerase chain reaction(dPCR);
copy number variant(CNV);
single nucleotide polymorphism(SNP);
cross-reactivity probe
- From:
Chinese Journal of Biochemistry and Molecular Biology
2025;41(6):895-902
- CountryChina
- Language:Chinese
-
Abstract:
Copy number variation encompassing SNP plays an important role in IVD and precision medi-cine.As the most commonly used method,FISH could not overcome the probe cross-reactivity which is common when to detect SNP.Here we developed a quantitative and qualitative method on copy number variation encompassing SNP.In this study,the rs76711854 was used as an example to establish a quanti-tative method by advantage of probe cross-reactivity.The fragment encompassing rs76711854 and its downstream to 9 514 bp were amplified by PCR.The allelic genotypes were verified by Sanger sequen-cing.Different probes with or without cross-reactivity to be used via quantitative real-time PCR and digit-al PCR.The different clusters(2D)and fluorescence intensity layers(1D)exist by adding probe with cross-reactivity.The A/G ratio measured by digital PCR is 2︰1,which is verified by probe targeting to the SNP.The copy-number variant exists in the 9kb-long fragment upstream to the SNP of prostate cancer cell line but not in human endometrial adenocarcinoma cell line Ishikawa.The data suggest that there is a multi-copy variation at this locus in DU145 cells.The method applied here is based on one single cross-reactivity probe via digital PCR.
