用套式PCR检测新生儿先天性巨细胞病毒感染

DETECTION OF HCMV CONGENITIAL INFECTION IN NEWBORNS BY NESTED-PCR

选取ＨＣＭＶＡＤ１６９株ＥｃｏＲＩＪ片段区的ＩＥＡ基因第四外显子为扩增靶序列，合成两对引物，外引物扩增２４２ｂｐ片段，内引物扩增１４６ｂｐ的片段，此方法很灵敏，可检测５－１０ｆｇＨＣＭｖＤＮＡ。我们随机采集５１名新生儿脐带血，应用ＮａＩ法提取全血ＤＮＡ为模板进行扩增，结果表明：３２名为ＨＣＭＶＤＮＡ阳性，以口腔粘液ＤＮＡ（４０名）为模板进行扩增，２４名为ＨＣＭＶＤＮＡ阳性。因此，套式ＰＣＲ可用于临床上快速准确地检测ＨＣＭＶ。

he primers sequences from the fourth exon of the HCMV IEA1 gene located in EcoRI Jfragment of strain AD 169 were synthesized.The first primer set was selecteed to amplify a 242bp fragment and the next primer set was nested within the first and amplified a 146 bp fragment. With the nested-PCR system it was possible to detect 5～10fg HCMV DNA, Fiftv-one cord bloodspecimens from newborns were collected randomly,treated with NaI and tested by the nested-PCR,32 HCMV DNA positive samples were found,while there were 24 HCMV DNA positivesamples among 40 mouth mucus specimens from newborns,The nested-PCR can be used forrapid, objective detection of HCMV DNA.