收稿日期: 2020-06-03
修回日期: 2020-08-06
网络出版日期: 2021-04-30
Thermal stability of diagnostic targets Plasmodium falciparum histidine rich protein Ⅱ and Plasmodium lactate dehydrogenase in rapid detection
Received date: 2020-06-03
Revised date: 2020-08-06
Online published: 2021-04-30
在新型冠状病毒肺炎流行期间,通常需将血样进行56 ℃病毒灭活处理后再行病原体检测。为了解56 ℃灭活对血样中疟原虫抗原稳定性的影响,本研究收集2017—2019年上海市疾病预防控制中心上报的疟疾患者血样71份,其中恶性疟血样38份、三日疟血样8份、卵形疟血样11份、间日疟血样14份。应用快速诊断检测（RDT）试剂盒对56 ℃孵育30 min前后血样中恶性疟原虫富组氨酸蛋白Ⅱ（PfHRPⅡ）和疟原虫乳酸脱氢酶（pLDH）的热稳定性进行测定。结果显示,热处理前T1检测线（检测目标PfHRPⅡ）呈阳性的38份恶性疟血样,热处理后35份仍呈T1阳性（92.11%,35/38, χ 2 = 3.123,P > 0.05）;热处理前T2检测线（检测目标pLDH）阳性的54份血样（26份恶性疟血样、6份三日疟血样、10份卵形疟血样和12份间日疟血样）,经热处理后T2均呈阴性（阳性率为0,0/54,χ 2 = 87.755, P < 0.01）。表明在56 ℃孵育30 min条件下,PfHRPⅡ的稳定性较好;pLDH不稳定,全部降解或失活。因此,应用RDT检测热处理后的血样,恶性疟血样的检测结果不受影响,但非恶性疟血样会被漏诊。
李美 , 涂宏 , 夏志贵 , 王真瑜 , 周何军 . 快速诊断靶蛋白恶性疟原虫富组氨酸蛋白Ⅱ和疟原虫乳酸脱氢酶的热稳定性[J]. 中国寄生虫学与寄生虫病杂志, 2021 , 39(2) : 245 -248 . DOI: 10.12140/j.issn.1000-7423.2021.02.022
During the COVID-19 epidemic, blood samples are usually processed at 56 ℃ to attenuate the virus before pathogen detection. 71 blood samples of malaria patients reported by Shanghai Center for Disease Control and Prevention in 2017—2019 were collected, including 38 with Plasmodium falciparum infection, 8 P. malariae, 11 P. ovale and 14 P. vivax. The effect of inactivation on the thermal stability of P. falciparum histidine rich protein Ⅱ (PfHRPⅡ) and Plasmodium lactate dehydrogenase (pLDH) in blood samples was assessed before and after incubation at 56 ℃ for 30 min using the rapid diagnostic test (RDT) kit. The results showed that among the 38 P. falciparum T1-positive (PfHRPⅡ) blood samples before heat treatment, 35 samples remained to be T1-positive (92.11%, 35/38, χ 2 = 3.123, P > 0.05) after heat treatment; while 54 blood samples (26 P. falciparum, 6 P. vivax, 10 P. ovale and 12 P. vivax) that were T2-positive (pLDH) before heat treatment turned to be T2-negative (positive rate 0, 0/54, χ 2 = 87.755, P < 0.01) after heat treatment. It was demonstrated that PfHRPⅡ is stable during incubation at 56 ℃ for 30 min, while pLDH is unstable and degraded or inactivated during the heating. Therefore, the detection results of P. falciparum will not be affected by RDT, but diagnosis of the parasites other than P. falciparum in blood samples may be missed.
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