Detection of three common G6PD gene mutations in Chinese individuals by probe melting curves.
OBJECTIVE
In order to develop and validate an assay for rapid detection of three common G6PD gene mutations in Chinese individuals.


METHODS
In this report we design two sets of primers and fluorescently labeled hybridization probes recognizing adjacent sequences in the amplicon; after annealing, the fluorophores were in resonance energy transfer, providing real-time monitoring of the amplication process. At the completion of PCR, fluorescence was monitored as the temperature increased through the Tm of the probe/product duplex, and a characteristic melting profile for each mutation was obtained. By using the fluorescence method and PCR/RE, a total of 57 samples obtained from two groups of G6PD-deficient individuals were studied.


RESULTS
A rapid method for detection of three common G6PD gene mutations in Chinese individuals by probe melting curves was developed. This method shows 100% accordance with the traditional method.


CONCLUSIONS
This fluorescent melting curve analysis is a simple, rapid, and effective method for clinical diagnosis and screening of G6PD deficiency.
Zhang
De-Tai
DT
Department of Laboratory Diagnosis, Union Hospital, Tong-ji Medical College, Hua-Zhong Science and Technology University, 1277# Jiefang Dadao, Wuhan 430022, PR China. detaizhangwh@163.com
Hu
Li-Hua
LH
Yang
Yu-Zheng
YZ
D002681:China; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D015342:DNA Probes; D005453:Fluorescence; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics*; D006801:Humans; D017403:In Situ Hybridization; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013696:Temperature; D044366:Transition Temperature
D017931:DNA Primers; D015342:DNA Probes; D005954:Glucosephosphate Dehydrogenase