荧光定量PCR在石蜡标本中检测新型隐球菌的应用价值  

The application value of fluorescence quantitative PCR in detecting Cryptococcus neoformans in paraffin-embedded specimens

作　　者：金晓燕 邹珏[1] 张倩倩[1] 董燕 胡慧娣[1] 沈丽华 JIN Xiaoyan;ZOU Jue;ZHANG Qianqian;DONG Yan;HU Huidi;SHEN Lihua(Department of Pathology,Nanjing Chest Hospital,Brain Hospital Affiliated to Nanjing Medical University,Nanjing,Jiangsu 210029,China)

机构地区：[1]南京市胸科医院/南京医科大学附属脑科医院病理科,江苏南京210029

出　　处：《临床肺科杂志》2025年第3期366-370,共5页Journal of Clinical Pulmonary Medicine

摘　　要：目的探索实时荧光定量逆转录PCR法(Reverse-transcription quantitative real-time polymerase chain reaction,RT-qPCR)在石蜡包埋肺组织中检出新型隐球菌的应用价值。方法采用传统染色方法(HE染色、六胺银染色、PAS染色)和RT-qPCR法对我院30例疑似新型隐球菌病病例进行检测,记录检测结果,用Kappa法进行一致性分析。结果四种方法都可以检出新型隐球菌,RT-qPCR法敏感度、特异度、阳性预测值、阴性预测值和确诊率(96.2%/100.0%/100.0%/80.0%/96.7%)均高于传统染色方法,HE染色(34.6%/75.0%/90.0%/15.0%/40.0%)、六胺银染色法(88.4%/50.0%/92.0%/40.0%/83.3%)和PAS染色法(80.7%/50.0%/91.3%/28.6%/76.7%),且与临床最终确诊的结果一致性较好(Kappa=0.870),而传统染色方法Kappa值均小于0.4。结论RT-qPCR法检测肺组织石蜡标本中新型隐球菌的敏感度高、特异度好,有望成为新型隐球菌快捷、稳定的检测手段之一。Objective To explore the application value of real-time fluorescent quantitative reverse transcription PCR(RT-qPCR)in the detection of Cryptococcus neoformans in paraffin-embedded lung tissues.Methods Traditional staining methods(HE staining,Gomori methenamine silver staining,PAS staining)and RT-qPCR were used to detect 30 cases suspected of cryptococcosis neoforme in our hospital.Record test results,using Kappa consistency analysis.Results Cryptococcus neoformans could be detected by all four methods.The sensitivity,specificity,positive predictive value(PPV),negative predictive value(NPV)and accuracy of the RT-qPCR(96.2%/100.0%/100.0%/80.0%/96.7%)method were higher than the traditional dyeing method such as HE staining(34.6%/75.0%/90.0%/15.0%/40.0%),Hexamine silver staining(88.4%/50.0%/92.0%/40.0%/83.3%),and PAS staining(80.7%/50%/91.3%/28.6%/76.7%),and had good consistent with the clinical diagnosis(Kappa=0.870),while the others was less than 0.4.Conclusion The RT-qPCR method has high sensitivity and specificity in detecting Cryptococcus neoforme in paraffin specimens of lung tissue,and it is expected to be one of the rapid and stable detection methods for Cryptococcus neoformans.

关 键 词：实时荧光定量逆转录PCR 新型隐球菌 石蜡包埋肺组织 

分 类 号：R519.4[医药卫生—内科学] R446.5[医药卫生—临床医学]