Application of fluorescence quantitative PCR in laboratory diagnosis of malaria

doi:10.12140/j.issn.1000-7423.2019.02.021

Abstract

Abstract:

Seventy-nine malaria samples were examined by microscopy, nested PCR and fluorescence quantitative PCR, and the detection rate was analyzed among the three methods. After combining clinical symptoms of the patients and results of the three methods, 74 of the 79 samples were confirmed to be malaria infection, with a positive rate of 93.7%（74/79）. Specifically, the detection rate of microscopy, nested PCR and fluorescence quantitative PCR were 82.3%（65/79）, 82.3%65/79） and 93.7%（74/79）, respectively. The concordance rate between microscopy and nested PCR was 78.4%（58/74）, and that between microscopy and fluorescence quantitative PCR was 63.5%（47/74）, between nested PCR and fluorescence quantitative PCR was 83.8%（62/74）. In addition, the detection rate of fluorescence quantitative PCR was significantly higher than that of the nested PCR（P < 0.05）. Compared with microscopy and nested PCR, the fluorescence quantitative PCR has higher sensitivity and positive detection rate, and is less time-consuming.

Key words: Malaria, Microscopy, Nested PCR, Fluorescence quantitative PCR

CLC Number:

R513.3

Su-hua LI, Jing LI, Li-jun GAO, Ya-lan ZHANG, Rui-min ZHOU, Dan QIAN, Cheng-yun YANG, Ying LIU, Yu-ling ZHAO, Hong-wei ZHANG. Application of fluorescence quantitative PCR in laboratory diagnosis of malaria[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2019, 37(2): 232-234.

Figures/Tables 3

References 13

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引物名称		引物序列（5′→3′）	产物长度/bp
第1轮	rPLU5	CCTGTTGTTGCCTTAAACTTC	1 600
	rPLU6	TCAAAGATTAAGCCATGCAAG
恶性疟原虫	rFAL1	TTAAACTGGTTTGGGAAAACC-AAATATATT	205
	rFAL2	ACACAATGAACTCAATCATGA-CTACCCGTC
间日疟原虫	rVIV1	CGCTTCTAGCTTAATCCACAT-AACTGATAC	120
	rVIV2	ACTTCCAAGCCGAAGCAAAG-AAAGTCCTTA
三日疟原虫	rMAL1	ATAACATAGTTGTACGTTAAG-AATAACCGC	144
	rMAL2	AAAATTCCCATGCATAAAAAA-TTATACAAA
卵形疟原虫	rOVA1	ATCTCTTTTGCTATTTTTTAGT-ATTGGAGA	800
	rOVA2	GGAAAGGACACATTAATTGT-ATCCTAGTG

样品编号	镜检	巢式PCR	荧光定量PCR
3	阴性	三日疟原虫	三日疟原虫
4	阴性	恶性疟原虫	恶性疟原虫
15	卵形疟原虫	阴性	卵形疟原虫
18	阴性	恶性疟原虫	恶性疟原虫
20	卵形疟原虫	阴性	卵形疟原虫
30	卵形疟原虫	阴性	卵形疟原虫
33	卵形疟原虫	阴性	卵形疟原虫
56	卵形疟原虫	阴性	卵形疟原虫
58	阴性	恶性疟原虫	恶性疟原虫
60	阴性	阴性	恶性疟原虫
62	阴性	阴性	恶性疟原虫
63	阴性	恶性疟原虫	恶性疟原虫
66	阴性	阴性	恶性疟原虫
74	阴性	恶性疟原虫	恶性疟原虫
75	卵形疟原虫	恶性疟原虫	卵形疟/恶性疟原虫
76	三日疟原虫	三日疟原虫	三日疟/恶性疟原虫
78	卵形疟原虫	阴性	卵形疟/恶性疟原虫
79	卵形疟原虫	卵形疟原虫	卵形疟/恶性疟原虫

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