Evaluation of detection method of ureaplasma urealyticum DNA and analysis of infection rate
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摘要:
目的对实时荧光定量PCR方法检测解脲脲原体（UU）核酸进行方法学评价。 方法依据CNAS-GL039性能评价要求文件及厂家声明，对本法重复性、准确度、测定下限和抗干扰能力进行验证；检测2018年1~12月我院门诊和住院患者UU标本共22 468例并统计分析。 结果该方法检测结果精密度均小于允许变异度；准确度与基因测序金标准符合率为100%；最低检测限为1×103 copies/mL；常见生殖道感染微生物不会产生交叉反应，4%的内源性红细胞对检测结果没有干扰；总UU阳性率为59.8%；女性UU检出率显著高于男性（χ2=525.613，P<0.01），女性<30>30岁年龄组差异具有统计学意义（P<0.01）。 结论qRT-PCR检测方法灵敏度高，重复性好，抗干扰力强，检测下限符合要求，可用于临床检测；本研究为UU感染诊断和治疗提供参考依据。 Abstract:ObjectiveTo evaluate the performance of real time quantitative PCR methods and analyze the infection status. MethodsAccording to the official CNAS-GL039 file and product specification, performance including precision, accuracy, lower limit of detection and anti-interference were investigated. Meanwhile, the samples from outpatients and inpatients in 2018 were collected and tested. The results were statistically analyzed. ResultsThe precision was within allowed coefficient of variation (CV), the accuracy was up to 100% by compared with the golden standard (gene sequencing). The lowest detection limit was 1×103 copies/mL. Cross-reaction between UU and common genital tract microorganisms were not present. The samples containing 4% of endogenous red blood cells did not interfere with test results. The overall positive rate of UU test was 59.8%. The detection rate in female was significantly higher than that in male (χ2=525.613, P<0 01="" among="" the="" positive="" samples="" of="" female="" patients="" the="" detection="" rate="" in="" 30="" years="" old="" was="" higher="" than="" that="" of="">30 years old (P<0.01). ConclusionThe detection method of qRT-PCR has a high detection sensitivity, good repeatability, strong anti-interference, and the lower limit of detection meets the requirements. It can be used for clinical testing. This study provides a reference basis for the diagnosis and treatment of UU infection. -
Key words:
- ureaplasma urealyticum /
- performance verification /
- epidemiological survey
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表 1 内源性红细胞实验结果
红细胞浓度 0% 2% 3.30% 4% Ct值（Mean±SD） 25.55±0.5 25.44±0.48 24.9±0.43 25.27±0.26 CV值（%） 1.9 1.89 1.73 1.03 
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表 2 UU总感染情况
性别 UU-DNA测定 检出数（n） 阳性率（%） 总数（n） 男 200 22.70 882 女 13 231 61.30* 21 586 *P<0.05 vs男性.
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表 3 不同年龄组男性UU-DNA检测结果
年龄组（岁） 阳性（n） 阳性率（%） 合计（n） 11~20 6 25.00 24 21~30 79 25.10 315 31~50 105 22.10 475 51~80 10 14.90 67
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表 4 不同年龄组女性UU-DNA检测结果
年龄组（岁） 阳性（n） 阳性率（%） 合计（n） 11~20 692 69.60 994 21~30 6 422 63.90 10 050 31~50 5 942 59.20 10 044 51~80 175 35.10 498
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