Thermal stability of diagnostic targets Plasmodium falciparum histidine rich protein Ⅱ and Plasmodium lactate dehydrogenase in rapid detection

doi:10.12140/j.issn.1000-7423.2021.02.022

Abstract

Abstract:

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.

Key words: Plasmodium antigens, Plasmodium falciparum histidine-rich protein Ⅱ, Plasmodium lactate dehydrogenase, Thermal stability

CLC Number:

R382.31

LI Mei, TU Hong, XIA Zhi-gui, WANG Zhen-yu, ZHOU He-jun. Thermal stability of diagnostic targets Plasmodium falciparum histidine rich protein Ⅱ and Plasmodium lactate dehydrogenase in rapid detection[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(2): 245-248.

Figures/Tables 1

References 20

[1]	Lei ZL, Wang LY. Control situation and primary task of key parasitic diseases in China[J]. Chin J Parasitol Parasit Dis, 2012,30(1):1-5. (in Chinese)
	( 雷正龙, 王立英. 全国重点寄生虫病防治形势与主要任务[J]. 中国寄生虫学与寄生虫病杂志, 2012,30(1):1-5.)
[2]	Zhang L, Feng J, Zhang SS, et al. The progress of national malaria elimination and epidemiological characteristics of malaria in China in 2017[J]. Chin J Parasitol Parasit Dis, 2018,36(3):201-209. (in Chinese)
	( 张丽, 丰俊, 张少森, 等. 2017年全国消除疟疾进展及疫情特征分析[J]. 中国寄生虫学与寄生虫病杂志, 2018,36(3):201-209.)
[3]	Zhang L, Feng J, Zhou SS. The progress of national malaria elimination and epidemiological characteristics of malaria in China in 2018[J]. 2019,37(3):241-247. (in Chinese)
	( 张丽, 丰俊, 周水森, 等. 2018年全国疟疾疫情特征及消除疟疾工作进展[J]. 中国寄生虫学与寄生虫病杂志, 2019,37(3):241-247.)
[4]	Zhang L, Feng J, Xia ZG. The progress of national malaria elimination and epidemiological characteristics of malaria in China in 2018[J]. 2020,38(2):133-138. (in Chinese)
	( 张丽, 丰俊, 夏志贵, 等. 2019年全国疟疾疫情特征及消除疟疾工作进展[J]. 中国寄生虫学与寄生虫病杂志, 2020,38(2):133-138.)
[5]	World Health Organization. WHO coronavirus disease (COVID-19) Dashboard[EB/OL]. (2021-04-12) [2021-04-12]. https://covid19.who.int/
[6]	Zou Y, Zheng YS, Cao J. Suggestions for standardizing the process of malaria diagnosis and treatment during the epidemic control of coronavirus disease (COVID-19)[J]. Chin J Parasitol Parasit Dis, 2020,38(1):1-4. (in Chinese)
	( 邹洋, 郑以山, 曹俊. 新型冠状病毒肺炎防控期间规范疟疾诊疗流程的建议[J]. 中国寄生虫学与寄生虫病杂志, 2020,38(1):1-4.)
[7]	Hussein M, Ali O, Homeida A, et al. Malaria and COVID-19: unmasking their ties[J]. Malar J, 2020,19(1):457. doi: 10.1186/s12936-020-03541-w
[8]	Chiodini J. COVID-19 and the impact on malaria[J]. Travel Med Infect Dis, 2020,35:101758. doi: S1477-8939(20)30245-3 pmid: 32479815
[9]	National Health Commission of the People’s Republic of China. The technical guidelines for novel coronavirus pneumonia diagnosis and treatment (version 7) [EB/OL]. (2020-03-04) [2020-06-03]. http://www.nhc.gov.cn/xcs/zhengcwj/202003/46c929-4a7dfe4cef80dc7f5912eb1989.shtml (in Chinese)
	( 中华人民共和国国家卫生健康委办公厅. 新型冠状病毒肺炎诊疗方案(试行第七版)[EB/OL]. (2020-03-04) [2020-06-03]. http://www.nhc.gov.cn/xcs/zhengcwj/202003/46c9294a7dfe4cef80-dc7f5912eb1989.shtml
[10]	WHO. Results of WHO product testing of malaria RDTs: round 7 (2015—2016) [M]. Geneva: WHO Press, 2017: 1-9.
[11]	WHO. Malaria Rapid Diagnostic Test Performance-Summary results of WHO malaria RDT product testing: Round 1-3 (2008—2011)[M]. Geneva: WHO Press, 2009: 1-16.
[12]	Jimenez A, Rees-Channer RR, Perera R, et al. Analytical sensitivity of current best-in-class malaria rapid diagnostic tests[J]. Malar J, 2017,16(1):128. doi: 10.1186/s12936-017-1780-5
[13]	Wang XY, Shi WZ. Protein chemistry and its application[M]. Beijing: China Textile Press, 2015: 45-64. (in Chinese)
	( 王雪燕, 师文钊. 蛋白质化学及其应用[M]. 北京: 中国纺织出版社, 2015: 45-64.)
[14]	Stillman JH, Somero GN. A comparative analysis of the evolutionary patterning and mechanistic bases of lactate dehydrogenase thermal stability in porcelain crabs, genus Petrolisthes[J]. J Exp Biol, 2001,204(Pt 4):767-776. pmid: 11171359
[15]	Wilson RJ McGregor IA Wilson RJ. The stability and fractionation of malarial antigens from the blood of Africans infected with Plasmodium falciparum[J]. Int J Parasitol, 1973,3(4):511-520. pmid: 4199255
[16]	Wu CP, Liu X, Li QW, et al. Structure and function of histidine-rich glycoprotein[J]. Chin J Biochem Mol Biol, 2012,28(1):1-8. (in Chinese)
	( 武彩萍, 刘欣, 李庆伟, 等. 富组氨酸糖蛋白(HRG)的结构与功能[J]. 中国生物化学与分子生物学报, 2012,28(1):1-8.)
[17]	Wilson RJ McGregor IA Hall PJ. Persistence and recurrence of S-antigen in Plasmodium falciparum infections in man[J]. Trans R Soc Trop Med Hyg, 1975,69(5/6):460-467. doi: 10.1016/0035-9203(75)90098-X
[18]	Dalrymple U, Arambepola R, Gething PW, et al. How long do rapid diagnostic tests remain positive after anti-malarial treatment?[J]. Malar J, 2018,17(1):228. doi: 10.1186/s12936-018-2371-9
[19]	Jiang L, Wang ZY, Ma XJ, et al. Comparative analysis of nucleotide sequences of lactate dehydrogenase (LDH) gene and LDH epitopes of Plasmodium vivax and Plasmodium falciparum[J]. Chin J Parasitol Parasit Dis, 2010,28(2):103-107. (in Chinese)
	( 江莉, 王真瑜, 马晓疆, 等. 间日疟原虫和恶性疟原虫乳酸脱氢酶基因的序列和重组抗原表位分析[J]. 中国寄生虫学与寄生虫病杂志, 2010,28(2):103-107.)
[20]	Liu ZY, Li XW, Zhou Y, et al. Impact of heating for virus inactivation on result of test of biochemical and inflammatory indexes[J]. Chin J Nosocomiol, 2020,30(10):1485-1489. (in Chinese)
	( 刘兆宇, 李学武, 周月, 等. 加热灭活病毒法对生化及炎症指标检测结果的影响[J]. 中华医院感染学杂志, 2020,30(10):1485-1489.)

血样	检测血样数	T1阳性血样数（占比/%）		χ²	P	T2阳性血样数（占比/%）		χ²	P	合计阳性血样数（占比/%）		χ²	P
血样	检测血样数	热处理前	热处理后	χ²	P	热处理前	热处理后	χ²	P	热处理前	热处理后	χ²	P
恶性疟	38	38(100)^a	35(92.1)	3.123	> 0.05	26(68.4)	0(0)	39.520	< 0.01	38(100)	35(92.1)	3.123	> 0.05
三日疟	8	0(0)	0(0)	-	-	6(6/8)	0(0)	9.600	< 0.01	6(6/8)	0(0)	9.600	< 0.01
卵形疟	11	0(0)	0(0)	-	-	10(10/11)	0(0)	7.619	< 0.01	10(10/11)	0(0)	7.619	< 0.01
间日疟	14	0(0)	0(0)	-	-	12(12/14)	0(0)	9.231	< 0.01	12(12/14)	0(0)	9.231	< 0.01
合计	71	38(50)	35(46.0)	0.237	> 0.05	54(71.0)	0(0)	83.755	< 0.01	66(86.8)	35(46.0)	16.189	< 0.01