Z Liu1, X Zhong, Z Li, X Xu. 1. Institute of Molecular Biology, First Military Medical Universty, Guangzhou,Guangdong,510515 P.R.China.
Abstract
OBJECTIVE: To establish a rapid and reliable PCR method for detecting a deletional hereditary persistence of fetal hemoglobin (HPFH) found in Chinese. METHODS: Based on some novel DNA sequences across the 3'breakpoint of a HPFH deletion observed in our laboratory, we designed a PCR amplification with three primers bridging the breakpoint. In this system, oligonucleotide primers have been chosen which allow specific identification of both normal and deletional chromosomes under identical condition in either the same or parallel PCR reactions. RESULTS: The expected two specific amplification bands were produced; one was 565bp in length and stood for the normal alleles,the other,37bp in length represented the mutant alleles of beta-globin gene cluster. This duplex system could directly genotype DNA samples bearing this type of HPFH deletion. CONCLUSION: This rapid and inexpensive method could be used as a routine method in the molecular screening of carriers and for the prenatal diagnosis of this disease.
OBJECTIVE: To establish a rapid and reliable PCR method for detecting a deletional hereditary persistence of fetal hemoglobin (HPFH) found in Chinese. METHODS: Based on some novel DNA sequences across the 3'breakpoint of a HPFH deletion observed in our laboratory, we designed a PCR amplification with three primers bridging the breakpoint. In this system, oligonucleotide primers have been chosen which allow specific identification of both normal and deletional chromosomes under identical condition in either the same or parallel PCR reactions. RESULTS: The expected two specific amplification bands were produced; one was 565bp in length and stood for the normal alleles,the other,37bp in length represented the mutant alleles of beta-globin gene cluster. This duplex system could directly genotype DNA samples bearing this type of HPFH deletion. CONCLUSION: This rapid and inexpensive method could be used as a routine method in the molecular screening of carriers and for the prenatal diagnosis of this disease.