西亞試劑：：建立一種人芳香二烷基磷酸酯酶(PON)基因叢等位基因快速分型法

目的　建立一種人芳香二烷基磷酸酯酶(PON)基因叢等位基因快速分型法。方法　在Multiplex-PCR-RELP基礎(chǔ)上，通過引物設(shè)計(jì)錯(cuò)配，在一體系中同時(shí)擴(kuò)增分別含PON1-192、PON1-55和PON2-31l位密碼子的DNA片段，當(dāng)?shù)任换驗(yàn)镻ON1-192R/PON1-55L/PON2-31lS時(shí)，PCR擴(kuò)增產(chǎn)物中均被引入唯一的限制性內(nèi)切酶HinfⅠ的識(shí)別位點(diǎn)G(上標(biāo) +)ANTC。PCR擴(kuò)增產(chǎn)物經(jīng)酶消化，聚丙烯酰胺凝膠電泳后，相互分開的不同組合的片段，確認(rèn)為不同的基因型組合，同時(shí)對(duì)3個(gè)位點(diǎn)進(jìn)行基因分型。結(jié)果　在檢測的80例健康個(gè)體中，等位基因頻率為PON1-192:Q46.9%.R53.1%:PON1-55:L95.6%，M4.4%；PON2-31l:S78.8%，C21.2%。結(jié)論　此方法快速分析3個(gè)位點(diǎn)的基因多態(tài)性，省時(shí)省力、節(jié)約經(jīng)費(fèi)，便于3個(gè)位點(diǎn)之間的連鎖分析，具有推廣價(jià)值。

Objective To establish an assay to identify paraoxonase gene cluster polymorphism. Methods Based on multiplex-PCR-RELP, we used mismatch primers that introduce an unique recognition site (Hinf Ⅰ) in PCR product in the presence of the R allele of PON1-192, L allele of PON1-55 and the S allele of PON-311 and then we simultaneously identified PON cluster polymorphism with electrophoretic band pattern specific for the variable combination through subsequent restriction analysis. Results The result shows that in 80 healthy individuals, the allelic frequency of PON1-192:Q46.9%, R 53.1%; PON1-55:L95.6% M4.4%; PON2-311:S78.8% C21.2% respectively. Conclusion This assay is an easy, economical and time-saving one characterized by detecting three polymorphism simultaneously.

丙烯醛肉桂醛桂皮醛4-硝基肉桂醛