不同分子检测技术对艾滋病患者合并寄生虫感染在真实世界的应用效果比较

Comparison on application effect of different molecular detection techniques in real world of acquired immune deficiency syndrome patients with parasitic infection

目的对住院艾滋病患者进行合并寄生虫感染的初步检验,比较不同DNA检测技术对艾滋病患者合并寄生虫感染在真实世界的应用效果。方法选择临床诊断为合并弓形虫感染或肺孢子虫感染的16例住院艾滋病患者作为研究对象,以临床诊断为“金标准”进行实验室分子检测。检测方法包括针对不同靶基因的荧光定量聚合酶链反应(qPCR)和环介导等温扩增技术(LAMP);临床样本包括脑脊液、血浆和支气管肺泡灌洗液(BALF);比较采用不同方法和临床样本的检测结果。结果采用Rep-529作为靶基因,LAMP与qPCR诊断艾滋病合并弓形虫感染的总检出率均为53.12%,差异无统计学意义(P>0.05)。对脑脊液样本进行LAMP-529-11和qPCR-529检测的检出率显著高于血浆样本〔90.62%(29/32)比15.62%(5/32),P<0.01〕。采用BALF样本临床诊断合并肺孢子虫肺炎的检出率显著高于血浆样本〔100.00%(32/32)比18.75%(6/32),P<0.01〕。LAMP-18S检测的检出率显著高于LAMP-Cob检测引物组〔65.62%(21/32)比46.87%(15/32),P<0.05〕。LAMP-18S与qPCR-mtSSU的总检出率比较差异无统计学意义〔65.62%(21/32)比53.12%(17/32),P>0.05〕。结论采用Rep-529作为靶基因对艾滋病患者合并弓形虫感染进行检测,qPCR和LAMP方法均可选,建议首选样本为脑脊液;肺孢子虫肺炎检测也可选qPCR和LAMP方法,建议首选样本为BALF。当选择LAMP方法时,建议首选针对18S的引物组。

Objective To preliminaryly detect the parasitic infection in hospitalized patients with acquired immune deficiency syndrome(AIDS)and compare the application effect of different DNA detection techniques on parasitic infection in AIDS patients in the real world.Methods The 16 patients with clinical diagnosis of AIDS and complicated with Toxoplasma gondii infection or Pneumocystis carinii infection were selected as research objects.The laboratory molecular testing was conducted using clinical diagnosis as"gold standard".The detection methods included fluorescence quantitative polymerase chain reaction(qPCR)and loop-mediated isothermal amplification(LAMP)for different target genes.The clinical samples included cerebrospinal fluid,plasma and bronchoalveolar lavage fluid(BALF).The detection results using different methods and clinical samples were compared.Results Using Rep-529 as target gene,the total detectable rates of LAMP and qPCR in diagnosing AIDS complicated with Toxoplasma gondii infection were both 53.12%,without statistically significant difference(P>0.05).The detectable rate of LAMP-529-11 and qPCR-529 method in cerebrospinal fluid samples was significantly higher than that in plasma samples[90.62%(29/32)vs.15.62%(5/32),P<0.01].The detectable rate of Pneumocystis carinii pneumonia in clinical diagnosis using BALF samples was significantly higher than that in plasma samples[100.00%(32/32)vs.18.75%(6/32),P<0.01].The detectable rate of LAMP-18S was significantly higher than that of LAMP-Cob detection primer group[65.62%(21/32)vs.46.87%(15/32),P<0.05].There was no statistically significant difference in the total detectable rate between LAMP-18S and qPCR-mtSSU[65.62%(21/32)vs.53.12%(17/32),P>0.05].Conclusions When Rep-529 is used as the target gene to detect Toxoplasma gondii infection in AIDS patients,both qPCR and LAMP methods are optional,and it is recommended that the first sample is cerebrospinal fluid.qPCR and LAMP methods could also be used for detecting Pneumocystis carinii pneumonia,and it is recommended to choose BALF as the preferred sample.When choosing the LAMP method,it is recommended to prioritize primer sets targeting 18S.