环介导等温扩增技术在检测妊娠期女性沙眼衣原体感染中的应用doi:10.3969/j.issn.1007-614x.2014.22.63摘要目的：评价酶联免疫吸附试验(ELISA)和环介导等温扩增方法(loop-mediatedisothermalamplification,LAMP)在检测妊娠期女性沙眼衣原体(ChlaniydiEietrachomatis,Ct)感染屮的应用价值。方法：根据Genebank公布的Cpn基因序列设计6条LAMP引物(2条内引物，2条外引物，2条环引物)，优化并建立LAMP检测体系。收治Ct感染患者80例，进行LAMP法检测CtDNA,同时用ELISA检测Ct特异性脂多糖抗原。结果:80例患者中,ELISA检测出阳性46例(57.5%),而LAMP检测出阳性56例(70.0%),两者比较差异无统计学意义(P>0.05)o80例患者中有51例服用过大环内酯类抗生素,ELTSA检测出阳性26例(50.9%),而LAMP检测出39例(76.5%),两者比较差异有统计学意义(P<0.05)o结论：LAMP检测泌尿生殖道Ct感染方法简便、快捷，尤其是对服用过抗生素的患儿也具有较高的检出率。关键词沙眼衣原体；环介导等温扩增技术；泌尿生殖道感染ApplicationofloopmediatedisothermalamplificationtechnologyinthedetectionofwomenchlamydiaetrachomatisinfectionduringpregnancyLiuWeil,LiuLiangzhuanl,WangHualil,HeLu2PathogenicBiologyResearchInstituteofNanhuaUniversity(HengyangCity,Hunan),4210011TheFirstHospitalAffiliatedtoNanhuaUniversity(HengyangCity,Hunan),4210012Abstractobjective：Toevaluatetheapplicationvalueofenzymelinkedimmunosorbentassay(ELISA)andloopmediatedisothermalamplification(LAMP)inthedetectionofwomenchlamydiaetrachomatis(Ct)infectionduringpregnancy.Methods：6LAMPprimers(2innerprimers,2outerprimers,2loopprimers)weredesignedaccordingtothesequenceofCpngenepublishedbyGenebank.LAMPdetectionsystemwasoptimizedandestablished.80caseswithCtwereselected.CtDNAwasdetectedbyLAMP・CtspecificfatpolysaccharideantigenwasdetectedbyELISA.Results：In80cases,ELISAdetectedthat46cases(57.5%)werepositive,whileLAMPdetectedthat56cases(70%)werepositive,andtherewasnostatisticalsignificance(P>0.05).In80cases,51casesweretreatedwithmacrolideantibiotics；ELISAdetectedthat26cases(50.9%)werepositive,whileLAMPdetectedthat39cases(76.5%)werepositive,andtherewasstatisticalsignificance(P<0.05).Conclusion：LAMPdetectionofurogenitalCtinfectionmethodissimpleandfast,especiallyithashigherdetectionrateforthechildrenwithtakingantibiotics.KeywordsChlamydiaetrachomatis；Loopmediatedisothermalamplificationtechnology；Urinogenitalinfection环介导等温扩增法(loop-mediatedisothermalamplification,LAMP)是一种敏感、特异、方便快捷的核酸扩增技术［1］。因其无需特殊设备，且具有PCR类似的灵敏度和特异性，H益受到临床重视［2］。沙眼衣原体(ChlamydieietmchonuHis,Ct)是一类严格细胞内寄生的病原体,不仅可以引起沙眼，还与人类各种泌尿生殖道疾病相关，如非淋菌性尿道炎、阴道炎、宫颈炎、子宫内膜炎等［3］；Ct还可通过母婴传染引起胎膜早破、早产、新生儿死亡等［4］。Ct感染初期无症状或症状较轻，常易被忽视而导致严重的并发症。本研究通过对比LAMP和酶联免疫吸附试验(ELISA)在检测妊娠期女性沙眼衣原体感染情况，分析LAMP在检测沙眼衣原体感染中的临床应用价值。资料与方法2010年1月-2012年1月收治沙眼衣原体感染患者80例，年龄25~39岁,平均(29±4.2)岁。所有患者均有不同程度的泌尿生殖道感染的临床症状，并且排除其他细菌感染性疾病。标本采集：先用无菌棉签拭去宫颈口周围的黏液及分泌物，将消毒的无菌棉拭子伸入宫颈口内2〜4cm,轻轻转动拭子，并停留10秒，退出拭子时，避免与阴道壁接触，拭子分别用于ELISA法和LAMP法检测。检测方法：①ELTSA：EIJSA检测试剂盒为Trinity公司产品(MicroTrakII,TrinityBiotech,Bray,Ireland),实验严格按照说明书操作:将处理好的样本加入96孔板中，96孔板中已包被有抗Ct脂多糖抗原(LPS)的兔抗体，如果样本中含有Ct的LP...