Performance comparison of three methods in detecting asymptomatic malaria infection on China-Myanmar border

doi:10.12140/j.issn.1000-7423.2020.02.004

Abstract

Abstract:

Objective To compare the performances of three methods in detecting asymptomatic malaria infection in residents, and assess the prevalence of asymptomatic malaria on China-Myanmar border.Methods Cross-sectional survey was carried out in Nabang town and Zhina town in Yingjiang county of Yunnan Province, China and a resettlement site in Laiza of Myanmar. Finger-prick blood samples were collected to prepare thick/thin blood smears and dried blood spots on filter paper. Malaria parasite infection was examined by light microscopy, real-time fluorescent PCR and ultrasensitive PCR (usPCR).Results A total of 387 blood samples were collected. Light microscopy found 6 participants with asymptomatic malaria infection (5 Plasmodium vivax infection and 1 P. falciparum infection), with a detection rate of 1.6%; real-time fluorescent PCR revealed 13 asymptomatic malaria infection cases (12 P. vivax infection; 1 P. falciparum infection) with a detection rate of 3.4%; and usPCR found 38 asymptomatic malaria infection cases(29 P. vivax infection and 9 P. falciparum infection) with a detection rate of 9.8%. Using light microscopy as the gold standard, the sensitivity and specificity of real-time fluorescent PCR were 98.2% and 100%, and those of usPCR were 91.6% and 100%, respectively. The usPCR results revealed that the detection rate of asymptomatic malaria infections was highest in Nabang town (17.1%, 22/129), followed by the Laiza resettlement site (10.0%, 11/110) and Zhina town (3.4%, 5/148), showing significant differences between the three sites (P < 0.05). Among the parasite species detected, P. vivax accounts for 76.3%, while P. falciparum for 23.7%. The detection rate was higher in females (10.7%, 23/215) than in males (8.7%, 15/172), but with no significant difference (P > 0.05). The age distribution showed that, the detection rate of asymptomatic infection was highest in the examinees aged 15-29 years(17.5%, 10/57), but there were no significant differences between the age groups.Conclusion Of the three methods compared for the ability in detection of asymptomatic malaria infection, the usPCR method presents higher detection rate than light microscopy and real-time fluorescent PCR. There are a certain proportion of asymptomatic malaria infection cases among the residents on China-Myanmar border.

Key words: Malaria, Asymptomatic malaria infection, PCR

CLC Number:

R531.3

Xiao-xiao WANG, Hui-hui XIAO, Fang HUANG, Shui-sen ZHOU. Performance comparison of three methods in detecting asymptomatic malaria infection on China-Myanmar border[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2020, 38(2): 152-158.

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方法Method	引物和探针名称 Primers and probes	序列（5′→3′） Sequences (5′→3′)
荧光PCR Fluorescent PCR	Pv-F	ATTGCTTTTGAGAGGTTTTGTTACTTT
	Pv-R	GCTGTAGTATTCAAACACAATGAACTCAA
	Pf-F	CGCTTCTAGCTTAATCCACATAACTG
	Pf-R	AATTTACTCAAAGTAACAAGGACTTCCAAG
超敏PCR Ultrasensitive PCR	Pf-F	CCGACTAGGTGTTGGATGAAAGTGTTAA
	Plas-R	AACCCAAAGACTTTGATTTCTCATAA
	Pf-P	/5Cy5/agcaatctaaaagtcacctcgaaagatgact/3BHQ_2/
	Pv-F	CCGACTAGGCTTTGGATGAAAGATTTTA
	Pv-P	/56-FAM/agcaatctaagaataaactccgaagagaaaattct/3BHQ_2/
	Actin-F	ACCGAGCGCGGCTACAG
	Actin-R	CTTAATGTCACGCACGATTTCC
	Actin-P	/VIC/ttcaccaccacggccgagc/MGB/

调查点 Spots	调查人数No. exam’d	显微镜观察Microscopic examination				荧光PCR Fluorescent PCR				超敏PCR Ultrasensitive PCR
调查点 Spots	调查人数No. exam’d	间日疟检出数 No. P. vivax infected	恶性疟检出数 No. P. falciparum infected	小计Subtotal	感染检出率/%Prevalence/%	间日疟检出数 No. P. vivax infected	恶性疟检出数 No. P. falciparum infected	小计Subtotal	感染检出率/%Prevalence/%	间日疟检出数 No. P. vivax infected	恶性疟检出数 No. P. falciparum infected	小计Subtotal	感染检出率/% Prevalence/%
那邦镇Nabang town	129	2	0	2	1.6	6	0	6	4.7	19	3	22	17.1
支那乡Zhina town	148	0	1	1	0.7	0	1	1	0.7	0	5	5	3.4
拉咱安置点Laiza resettlement site	110	3	0	3	2.7	6	0	6	5.5	10	1	11	10.0
合计Total	387	5	1	6	1.6	12	1	13	3.4	29	9	38	9.8

方法和结果Methods and results		显微镜观察Microscopic examination		小计Subtotal	灵敏度/%Sensitivity/%	特异度/%Specificity/%
方法和结果Methods and results		阳性数No. positive	阴性数No. negative	小计Subtotal	灵敏度/%Sensitivity/%	特异度/%Specificity/%
荧光PCR Fluorescent PCR	阳性数No. positive	6	7	13
荧光PCR Fluorescent PCR	阴性数No. negative	0	374	374	100	98.2
	小计Subtotal	6	381	387
超敏PCR Ultrasensitive PCR	阳性数No. positive	6	32	38
	阴性数No. negative	0	349	349	100	91.6
	小计Subtotal	6	381	387

特征 Feature	感染检出例数No. infected	总例数 Total	比例/% Percent
性别Gender
男Male	15	172	8.7
女Female	23	215	10.7
年龄组Age group
0~4	1	140	0.7
5~14	7	56	12.5
15~29	10	57	17.5
30~44	10	66	15.2
45~60	8	49	16.3
> 60	2	19	2/19
虫种Species
间日疟原虫P. vivax	29	387	7.5
恶性疟原虫P. falciparum	9	387	2.3