HOMEPRODUCTSCOMPANYCONTACTFAQResearchDictionaryPharmaSign Up FREE or Login

A melting curve analysis--based PCR assay for one-step genotyping of β-thalassemia mutations a multicenter validation.

Abstract
The increasing number of disease-causing mutations demands a simple, direct, and cost-effective diagnostic genotyping technique capable of detecting multiple mutations. This study validated the efficacy of a novel melting curve analysis-based genotyping assay (MeltPro HBB assay) for 24 β-thalassemia mutations in the Chinese population. The diagnostic potential of this assay was evaluated in 1022 pretyped genomic DNA samples, including 909 clinical cases of β-thalassemia minor or major, using a double-blind analysis in a multicenter validation study. Reproducibility of the assay was 100%, and the limit of detection was 10 pg per reaction. All 24 β-thalassemia mutations were accurately genotyped, and β-thalassemia genotypes were correctly determined in all 1022 samples, yielding overall sensitivity and specificity of 100%. The concordance rate was 99.4% between this assay and the reference method. It was concluded that the MeltPro HBB assay is useful for reliable genotyping of multiple β-thalassemia mutations in clinical settings and may have potential as a versatile method for rapid genotyping of known mutations because of its high throughput, accuracy, ease of use, and low cost.
AuthorsFu Xiong, Qiuying Huang, Xiaoyun Chen, Yuqiu Zhou, Xinhua Zhang, Ren Cai, Yajun Chen, Jiansheng Xie, Shanwei Feng, Xiaofeng Wei, Qizhi Xiao, Tianlang Zhang, Shiqiang Luo, Xuehuang Yang, Ying Hao, Yanxia Qu, Qingge Li, Xiangmin Xu
JournalThe Journal of molecular diagnostics : JMD (J Mol Diagn) Vol. 13 Issue 4 Pg. 427-35 (Jul 2011) ISSN: 1943-7811 [Electronic] United States
PMID21704277 (Publication Type: Journal Article, Multicenter Study, Research Support, Non-U.S. Gov't, Validation Study)
CopyrightCopyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
Chemical References
  • Beta-Globulins
Topics
  • Beta-Globulins (genetics)
  • DNA Mutational Analysis (methods)
  • Double-Blind Method
  • Humans
  • Limit of Detection
  • Molecular Diagnostic Techniques
  • Polymerase Chain Reaction
  • Reproducibility of Results
  • Transition Temperature
  • beta-Thalassemia (diagnosis, genetics)

Join CureHunter, for free Research Interface BASIC access!

Take advantage of free CureHunter research engine access to explore the best drug and treatment options for any disease. Find out why thousands of doctors, pharma researchers and patient activists around the world use CureHunter every day.
Realize the full power of the drug-disease research graph!


Choose Username:
Email:
Password:
Verify Password:
Enter Code Shown: