Establishment of a system for rapid detection of JAK2 V617F mutation in myeloproliferative diseases.
10.3760/cma.j.issn.1003-9406.2019.10.007
- VernacularTitle:骨髓增生疾病JAK2 V617F突变快速检测体系的建立
- Author:
Qiang ZHANG
1
,
2
;
Yijia ZHANG
;
Xiangmin XU
;
Wanjun ZHOU
;
Xin FAN
Author Information
1. Central Laboratory for Inborn Error of Metabolism, Maternal and Child Health Care Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi 530003, China. fanxin602@
2. com.
- Publication Type:Journal Article
- MeSH:
Alleles;
DNA Mutational Analysis;
Double-Blind Method;
Humans;
Janus Kinase 2;
genetics;
Mutation;
Myeloproliferative Disorders;
genetics;
Real-Time Polymerase Chain Reaction;
Sensitivity and Specificity
- From:
Chinese Journal of Medical Genetics
2019;36(10):980-984
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To develop a system for rapid detection of JAK2 V617F mutation among patients with myeloproliferative diseases.
METHODS:Specific primers and TagMan probes were designed for the mutant and wild type alleles based on the principle of real-time PCR. A complete system including the method for detection and product for quality control were established through the evaluation of sensitivity and accuracy of the method, double-blind trial, and preparation of negative and positive controls through site-directed mutagenesis and molecular cloning.
RESULTS:A system for rapid detection of the JAK V617F mutation has been developed. Compared with Sanger sequencing, the sensitivity and specificity of the method have both reached 100%. Meanwhile, 1000 normal samples and 1 case with the JAK2 V617F mutation were detected, which gave a population rate of 1‰.
CONCLUSION:The system was fast, accurate, cheap, high throughput, and easy to use. It can be utilized as a routine test. Although the JAK2 V617F mutation is rare in the population, it should be screened among myeloproliferative neoplasm patients.
