解脲脲原体DNA检测方法评价及人群感染情况调查

宗曾艳; 熊丹; 汤花梅; 王萌萌; 孔凡虹; 阚丽娟; 张水兰; 张秀明

doi:10.12122/j.issn.1674-4500.2019.04.28

解脲脲原体DNA检测方法评价及人群感染情况调查

doi: 10.12122/j.issn.1674-4500.2019.04.28

宗曾艳^1,2,,
熊丹^1,2,
汤花梅²,
王萌萌^1,2,
孔凡虹²,
阚丽娟²,
张水兰²,
张秀明^1,2, ,

1.
安徽理工大学，安徽淮南 232000
2.
深圳市罗湖区人民医院检验科，广东深圳 440301

详细信息

作者简介:
宗曾艳，硕士，E-mail：1445385411@qq.com

通讯作者:
张秀明，硕士生导师，主任技师，E-mail：zxm0760@163.com

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- 被引次数: 0
出版历程
- 收稿日期: 2019-09-18
- 刊出日期: 2019-12-01

Evaluation of detection method of ureaplasma urealyticum DNA and analysis of infection rate

1.
Anhui University of Science and Technology, Huainan 232000, China
2.
Department of Laboratory Medicine, Luohu District People's Hospital, Shenzhen 440301, China

摘要

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。
- 解脲脲原体 /
- 性能验证 /
- 流行病学调查
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.
- ureaplasma urealyticum /
- performance verification /
- epidemiological survey

HTML全文

图 1 UU-DNA测序实验凝胶电泳结果

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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
性别	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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