PCR荧光法检测人类HLA-B27基因的临床应用

The clinical application of fluorescent PCR method in human HLA-B27 detection

目的依据ISO15189规定评价PCR荧光法检测人类HLA-B27基因在临床中的实际应用性,涉及阴阳性符合率、精密度、灵敏度和抗干扰能力。方法依据ISO15189认可要求对试剂盒的准确度(与金标准比较)、精密度、检测下限和抗干扰能力进行性能评价。结果将40例样本检测结果与测序结果进行比对,符合率100%。低浓度样本批内精密度为1.13%、批间精密度为1.00%,高浓度样本批内精密度为0.98%、批间精密度为0.88%,均符合厂家要求CV≤5%。检测下限即检测最低线CT值为38,重复检测20次,检出率为100%。抗干扰能力检测结果与原样本检测结果阴阳性100%符合。结论 PCR荧光法检测人类HLA-B27基因结果稳定、准确、可靠,满足ISO15189认可要求,可用于临床人类HLA-B27基因的检测。

Objective To evaluate the method of fluorescent PCR to detect human HLA-infection among B27-carriers based on ISO15189 regulation,involving the masculine coincidence rate,precision,sensitivity and anti-interference ability.Methods Performance verification was conducted according to the ISO15189 accreditation,including accuracy(comparing with gold standard),precision,limit of lowest detection and anti-interference ability.Results The samples of 40 cases were tested positive results for 30 cases,negative for 10 cases,coincidence rate was 100%.A precision of 1.13%in low concentration sample batch,batch precision was 1.00%;a precision of 0.98%in high concentration sample batch and batch precision was 0.88%,and conformed to the requirements of the manufacturer CV 5%or less.Detection limit for the lowest line CT value was 38,repeated testing 20 times,and detection rate was 100%.The detection results of anti-interference ability were 100%in line with the positive and negative results of the original samples.Conclusion PCR fluorescence detecting human HLA-infections among B27-carriers result is stable,accurate and reliable,and it can be used to'clinical detection of human HLA-infections among B27-carriers'.