3种疟疾检测方法的应用分析

摘要/Abstract

摘要：

目的分析疟原虫镜检、抗原检测（快速诊断试验,RDT）和核酸检测（PCR）3种方法在疟疾病例检测应用中的效果。方法对2012-2015年上海市送检的疟疾病例和疑似疟疾病例血样的检测结果进行回顾性分析,以确诊病例为金标准,比较镜检、RDT和PCR 等3种方法的诊断指标、虫种特异性鉴定能力和成本效益。不同检测方法的两两比较用χ²检验。结果 3种方法平行分析血样212份,结果显示,综合判定为阳性的167份,占78.8%,阴性45份,占21.2%。阳性血样中,120份为恶性疟原虫（Plasmodium falciparum）感染,占71.9%;22份为间日疟原虫（P. vivax）感染,占13.2%;17份为卵形疟原虫（P. ovale）感染,占10.2%;6份为三日疟原虫（P. malariae）感染,占3.6%;2份为混合感染,占1.2%。3种检测方法的诊断效率以PCR最高,为96.2%（204/212）,RDT次之,为93.2%（192/206）,镜检最低,为88.2%（187/212）;镜检与RDT和PCR比较,差异具有统计学意义（P < 0.05）;RDT与PCR比较,差异无统计学意义（P > 0.05）。与确诊病例比较,3种方法的总符合率以PCR最高,为95.3%（202/212）,RDT次之,为93.2%（192/206）,镜检最低,为88.2%（187/212）;镜检与PCR比较,差异有统计学意义（P < 0.05）。对非恶性疟原虫的3种疟原虫的种特异性鉴定能力,以PCR最高,为95.6%（43/45）,镜检次之,为91.1%（41/45）,RDT最低,为68.9%（31/45）;PCR与RDT比较,差异有统计学意义（P < 0.05）;PCR与镜检比较,差异无统计学意义（P > 0.05）。对恶性疟原虫单一虫种的鉴定能力,以RDT最优,为100%（116/116）;PCR次之,为93.3%（112/120）;镜检最低,为84.2%（101/120）。根据是否纳入诊断标准、对仪器设备和技术的要求、诊断效能、检测的成本耗时、应用培训以及技术推广等14项指标综合评测显示,RDT得分为37分（满分42分）,成本效益最高,镜检和PCR得分分别为26和27分。结论在以输入性恶性疟为主的疫情形势下,3种检测方法比较,检测效果以PCR和RDT较高,种特异性鉴定能力以PCR和镜检较好,成本效益以RDT最佳。

关键词: 上海市, 疟疾, 镜检, 快速诊断试验, PCR

Abstract:

Objective To test the usage of microscopic examination, antigen detection（rapid dignostic test, RDT） and nucleic acid test（PCR） for detection of malaria cases. Methods The blood test results for malaria and suspected malaria cases during 2012-2015 were retrospectively reviewed. Taking the confirmed cases as a gold standard, the three methods were compared in aspects of diagnosis indices, specificity of identification species, and cost effectiveness. Results A total of 212 samples were included, each analyzed with the three methods. Based on the results of the three tests, 167 （78.8%） were determined to be positive for malaria, and 45 negative （21.2%）. Of the positive samples, 120 （71.9%） were infected with Plasmodium falciparum, 22 （13.2%） with P. vivax, 17（10.2%） with P. ovale, 6 （3.6%） with P. malariae, and 2 （1.2%） with mixed infections. The method of PCR had the highest diagnostic efficiency （96.2%, 204/212）, followed by RDT （93.2%, 192/206; P > 0.05 vs. PCR） and the microscopic method （88.2%, 187/212; P < 0.05 vs. RDT and PCR）. Similarly, the PCR method had the highest overall coincidence rate to the confirmed cases （95.3%, 202/212）, followed by RDT （93.2%, 192/206） and microscopy （88.2%, 187/212; P < 0.05 vs. PCR）. As to the identification specificity among species, the PCR method（95.6%, 43/45） was superior to microscopy （91.1%, 41/45; P > 0.05 vs. PCR） and RDT （68.9%, 31/45; P < 0.05 vs. PCR）. As to the identification of a particular species （P. falciparum）, RDT performed best （100%, 116/116）, followed by PCR （93.3%, 112/120） and microscopy （84.2%, 101/120）. Based on the comprehensive evaluation on 14 indicators including if it is a diagnostic criterion, equipment and technical requirement, diagnostic performance, time cost, and the need of technical training and promotion, we found that the RDT method had the highest score（37 of 42）, while microscopy and PCR were scored 26 and 27, respectively. Conclusion Under the falciparum malaria-dominated epidemiological situation, PCR and RDT show a higher detection efficiency, PCR and microscopy perform better in species identification, and RDT has the highest cost-effectiveness.

Key words: Shanghai, Malaria, Microscopy, Rapid diagnostic test, PCR

中图分类号:

R531.3

江莉, 王真瑜, 张耀光, 朱民, 张小萍, 马晓疆, 何燕艳, 朱倩, 蒋守富, 蔡黎. 3种疟疾检测方法的应用分析[J]. 中国寄生虫学与寄生虫病杂志, 2017, 35(1): 53-58.

Li JIANG, Zhen-yu WANG, Yao-guang ZHANG, Min ZHU, Xiao-ping ZHANG, Xiao-jiang MA, Yan-yan HE, Qian ZHU, Shou-fu JIANG, Li CAI. Analysis on the application of three methods for malaria diagnosis[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2017, 35(1): 53-58.

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参考文献 17

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虫种 Species	确诊病例数 No. confirmed cases	镜检 Microscopic examination		抗原检测 RDT		核酸检测 PCR
虫种 Species	确诊病例数 No. confirmed cases	检出数 No. detected	符合率/% Coincidence rate/%	检出数 No. detected	符合率/% Coincidence rate/%	检出数 No. detected	符合率/% Coincidence rate/%
恶性疟原虫 P. falciparum	120	101	84.2	116	100^b	112	93.3
间日疟原虫 P. vivax	22	21				22
卵形疟原虫 P. ovale	17	16	91.1	31	68.9	15	95.6
三日疟原虫 P. malariae	6	4				6
混合感染 Mixed infection	2	0	-	0	-	2	-
未分型Unclassified	0	0	-	0	-	2	-
阴性 Negative	45	70	-	59	-	53	-
未检测^a Untested^a	0	0	-	6	-	0	-
总计 Total	212	212	-	212	-	212	-

指标 Indicators	镜检Microscopic examination	得分Score	PCR	得分 Score	RDT	得分 Score
纳入诊断标准 Diagnostic criteria	是Yes	3	是Yes	3	是Yes	3
专用仪器设备 Need of special equipment	需要Yes	1	需要Yes	1	不需要No	3
技术要求 Technical requirement	高High	1	高High	1	低Low	3
经验对检测结果影响 Effect of experience on results	大Significant	1	中Medium	2	小Little	3
确诊现患感染 Diagnosis of active infection	是Yes	3	是Yes	3	是Yes	3
鉴定虫种 Species identified	4种4 species	3	4种4 species	3	1~2种1-2 species	2
敏感性Sensitivity	低Low	1	高High	3	中Medium	2
特异性Specificity	中Medium	2	高High	3	低Low	1
检测耗时Time-cost	1.5 h	2	3~4 h	1	10~20 min	3
试剂成本Reagent cost	低Low	3	高High	1	中低Medium	2
培训要求Training	高High	1	中Medium	2	低Low	3
现场检测Field test	否No	1	否No	1	是Yes	3
采样难度Sampling	易Easy	3	难Hard	1	易Easy	3
技术推广Application	难Hard	1	中等Medium	2	易Easy	3
得分合计（百分制评分） Score （percent score）	26（62）		27（64）		37（88）