目的 对2015—2021年辽宁省各地市上报的疟疾病例样本,在省级参比实验室进行检测复核,并根据实验结果评估各地市疟疾诊断参比实验室的疟疾诊断能力,为进一步提升各地市疾病预防控制（疾控）中心疟疾诊断能力提供参考依据。方法 对上报的血涂片样本进行镜检复核,疟疾病例的抗凝血样本进行巢式PCR和荧光定量PCR（qPCR）检测,采用χ2检验和Fisher确切概率法对各组间检出率、检出率与上报阳性率间差异进行统计学分析。结果 2015—2021年累计复核380份疟疾病例样本,其中恶性疟阳性样本270份,间日疟20份,三日疟11份,卵形疟34份,阴性45份。上报病例样本主要集中在沈阳和大连市,共278份,占全省病例总数的73.16%。对各地市实验室检测的复核结果分析显示,沈阳市检测符合率为95.36%,大连市为92.97%,全省实验室平均符合率为90.13%。比较镜检和巢式PCR 2种方法的结果,巢式PCR检测阳性率为87.63%,血涂片镜检阳性率为74.47%,2种方法检出率差异有统计学意义（χ2=46.296,P<0.001）。结论 辽宁省各地市疾控中心在血涂片镜检和核酸检测能力虽然逐步提高,但检测水平不均衡,仍然有较大的提升空间,需继续加强实验室人员检测能力培训,稳固实验室检测队伍人员应有的检测水平,做到送检样本的及时精准检测与反馈,巩固疟疾消除成果。
Objective To assess the diagnostic capacities for malaria of prefecture-level malaria diagnostic reference laboratories of Liaoning Province, China by checking the samples from malaria cases provided by each city from 2015 to 2021 at the provincial reference laboratory, so as to provide a basis for further improving the malaria diagnostic capacity of the centers for disease control and prevention in each city. Methods The blood smear samples were checked under a microscope, and the anticoagulated blood samples from malaria cases were tested using nested PCR and quantitative real time PCR. The Chi-square test and Fisher's exact test were used to compare the detection rate between groups, and to compare the detection rate and reported positive rate. Results A total of 380 samples from malaria cases were checked from 2015 to 2021, in which 270 were positive for Plasmodium falciparum, 20 were positive for P. vivax, 11 were positive for P. malariae, 34 were positive for P. ovale, and 45 were negative. The samples were mainly from Shenyang and Dalian, with a total of 278 samples, accounting for 73.16% of all the samples of the province. The check results showed that the coincidence rate was 95.36% for Shenyang and 92.97% for Dalian, and the average coincidence rate of all the laboratories was 90.13%. The positive rate by nested PCR was 87.63%, and the positive rate by blood smear microscopy was 74.47%, which were significantly different (χ2=46.296, P<0.001). Conclusions Although progress has been made in the capabilities of blood microscopy and nucleic acid detection of prefecture-level centers for disease control and prevention of Liaoning Province, their detection levels are imbalanced, and there is still a large room for improvement. In order to consolidate the malaria elimination achievements, it is necessary to continue to strengthen laboratory personnel training on testing ability, stabilize the testing level, and make sure timely and accurate detection and feedback.
[1] Li S, Zhang YH, Wang YY, et al. Hot spots review of global infectious diseases in 2016[J]. Infect Dis Inform, 2017, 30（1）: 1-7. DOI:10.3969/j.issn.1007-8134.2017.01.003.（in Chinese） 李硕, 张云辉, 王永怡, 等. 2016年全球传染病热点回顾[J]. 传染病信息, 2017, 30（1）：1-7. DOI:10.3969/j.issn.1007-8134.2017.01.003.
[2] Qi XQ. Malaria control manual[M]. 3rd ed. Beijing: People's Medical Publishing House, 2007: 78-79.（in Chinese） 齐小秋. 疟疾防治手册[M]. 3版. 北京: 人民卫生出版社, 2007: 78-79.
[3] Zhang YG, Jiang L, Wang ZY, et al. Analysis of malaria detection capability of laboratories in the districts of Shanghai during 2017-2019[J]. Chin J Parasitol Parasit Dis, 2021, 39（3）: 337-342. DOI:10.12140/j.issn.1000-7423.2021.03.007.（in Chinese） 张耀光, 江莉, 王真瑜, 等. 2017—2019年上海市各区疟疾实验室检测能力分析[J]. 中国寄生虫学与寄生虫病杂志, 2021, 39（3）: 337-342. DOI:10.12140/j.issn.1000-7423.2021.03.007.
[4] World Health Organization. World malaria report 2020[R]. Geneva: WHO, 2020: 18, 20.
[5] Gao Q. Risk and countermeasures of re-establishment after malaria elimination in China[J]. China Trop Med, 2024, 24（1）: 1-5. DOI:10.13604/j.cnki.46-1064/r.2024.01.01.（in Chinese） 高琪. 当前疟疾形势与我国消除疟疾后的再传播风险与对策[J]. 中国热带医学, 2024, 24（1）: 1-5. DOI:10.13604/j.cnki.46-1064/r.2024.01.01.
[6] Zhu GD, Gao Q, Cao J. Challenges and countermeasures in prevention of re-establishment of imported malaria in China[J]. Chin J Schisto Control, 2021, 33（1）: 7-9, 21. DOI:10.16250/j.32.1374.2021008.（in Chinese） 朱国鼎, 高琪, 曹俊. 中国防止疟疾输入再传播面临的挑战和应对策略[J]. 中国血吸虫病防治杂志, 2021, 33（1）: 7-9, 21. DOI:10.16250/j.32.1374.2021008.
[7] Feng J, Zhang L, Huang F, et al. Ready for malaria elimination: Zero indigenous case reported in the People's Republic of China[J]. Malar J, 2018, 17（1）: 315. DOI:10.1186/s12936-018-2444-9.
[8] Zhu GD, Cao J. Progress and challenges of global malaria elimination[J]. Chin J Schisto Control, 2019, 31（1）: 19-22, 52. DOI:10.16250/j.32.1374.2019006.(in Chinese)朱国鼎, 曹俊. 全球消除疟疾进展及面临的挑战[J]. 中国血吸虫病防治杂志, 2019, 31（1）:19-22, 52. DOI:10.16250/j.32.1374.2019006.
[9] Feng J, Xiao HH, Xia ZG, et al. Analysis of malaria epidemiological characteristics in the People's Republic of China, 2004-2013.[J]. Am J Trop Med Hyg, 2015, 93（2）: 293-299. DOI:10.4269/ajtmh.14-0733.
[10] Feng J, Zhang L, Xia ZG, et al. Malaria-free certification in China: Achievements and lessons learned from the National Malaria Elimination Programme[J]. Zoonoses, 2021, 1（1）：3-6. DOI:10.15212/ZOONOSES-2021-1002.
[11] Wang B, Mao LL, Wang ZJ, et al. Imported oval malaria in Liaoning Province, China, in 2017: A laboratory diagnosis of 5 cases[J]. Chin J Vector Biol Control, 2019, 30（1）: 80-83. DOI:10.11853/jissn.1003.8280.2019.01.019.（in Chinese） 王博, 毛玲玲, 王子江, 等. 辽宁省2017年5例输入性卵形疟疾的实验室诊断[J]. 中国媒介生物学及控制杂志, 2019, 30（1）: 80-83. DOI:10.11853/j.issn.1003.8280.2019.01.019.
[12] World Health Organization. Guidelines for the treatment of malaria[M]. 2nd ed. Geneva: WHO, 2010: 35-36.
[13] Snounou G, Viriyakosol S, Zhu XP, et al. High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction[J]. Mol Biochem Parasitol, 1993, 61（2）: 315-320. DOI:10.1016/0166-6851(93)90077-b.
[14] Snounou G, Viriyakosol S, Jarra W, et al. Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections[J]. Mol Biochem Parasitol, 1993, 58（2）：283-292. DOI:10.1016/0166-6851(93)90050-8.
[15] Singh B, Bobogare A, Cox-Singh J, et al. A genus-and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies[J]. Am J Trop Med Hyg, 1999, 60（4）: 687-692. DOI:10.4269/ajtmh.1999.60.687.
[16] Teng C, Lei L, Sun YW, et al. Prevalence of malaria in Liaoning Province, 2008 to 2013[J]. Chin J Schisto Control, 2014, 26（2）: 200-202. DOI:10.16250/j.32.1374.2014.02.005.（in Chinese） 滕聪, 雷露, 孙英伟, 等. 2008—2013年辽宁省疟疾流行病学分析[J]. 中国血吸虫病防治杂志, 2014, 26（2）：200-202. DOI:10.16250/j.32.1374.2014.02.005.
[17] Li M, Wang ZY, Zhang T, et al. Exploration of using one-step reverse transcription PCR in detection of four species of human malaria parasites[J]. Chin J Parasitol Parasit Dis, 2016, 34（6）: 500-505.（in Chinese） 李美, 王真瑜, 张淘, 等. 一步反转录PCR技术在检测4种人疟原虫中的初步应用[J]. 中国寄生虫学与寄生虫病杂志, 2016, 34（6）: 500-505.
