Establishment and evaluation of colloid gold-labeled immunochromatographic test strip for rapid detection of antibody against Necator americanus

Abstract

Abstract:

Objective To establish and evaluate colloid gold-labeled immunochromatographic test strip for rapid and convenient detection of antibody against Necator americanus. Methods Adult worms of N. americanus was ground in liquid nitrogen, immersed in membrane protein surfactant, followed by repeated freezing and thawing in liquid nitrogen and ammonium sulfate precipitation, to extract soluble antigen. The colloid gold-labeled immunochromatographic test strip was established using the soluble antigen as a coating antigen and the colloidal gold-labeled G protein as the detective probe. This test strip was used to examine sera from patients infected with N. americanus, Ascaris lumbricoides, Trichuris trichiura, Schistosoma japonicum and Toxoplasma gondii and sera from healthy participants, to evaluate the sensitivity and specificity of the assay, accompanied by ELISA as a parallel control to evaluate the detection efficacy. Results The colloid gold-labeled immunochromatographic test strip for detecting antibody against N. americanus was established. This test was performed to examine sera from patients infected with N. americanus (95 samples), A. lumbricoides (10 samples), T. trichiura (11 samples), S. japonicum (10 samples) and T. gondii (10 samples) and 74 serum samples from healthy participants, accompanied by ELISA as a parallel. The sensitivity of the colloid gold-labeled immunochromatographic strip test and ELISA in detecting N. americanus was 88.4% （84/95) and 90.5% （86/95), respectively, with false positive rates of 4.1% （3/74） and 6.8% （5/74） in 74 healthy serum samples, with rates of cross reaction with ascariasis being 2/10 and 4/10, rates of cross reaction with trichuriasis being 1/11 and 4/11. The strip test had no cross reaction with schistosomiasis or toxoplasmosis, while ELISA had a cross reaction rate of 1/10 with schistosomiasis but no cross reaction with toxoplasmosis. The specificity of the strip test and ELISA was 94.9% (109/115) and 88.7% (102/115), and the diagnostic efficacy was 91.9% and 89.5%, respectively. There was no significant difference in sensitivity （P > 0.05） or specificity （P > 0.05） between the strip test and ELISA. Conclusion The colloid gold-labeled immunochromatographic test strip shows relatively high sensitivity and specificity in diagnosis of ancylostomiasis.

Key words: Ancylostomiasis, Necator americanus, Soluble antigen, Immunochromatographic strip, Diagnosis

CLC Number:

R383.132

Feng SHI, Yi YANG, Jun-yun WANG, Yue-tao YANG, Chun-hua GAO. Establishment and evaluation of colloid gold-labeled immunochromatographic test strip for rapid detection of antibody against Necator americanus[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2018, 36(2): 119-123.

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血清 Serum	检测数 No. detected	阳性数No. positives
血清 Serum	检测数 No. detected	试条法 Immunochromatographic strip test	ELISA法 Enzyme-linked immunosorbent assay
钩虫感染 Infected with Necator americanus	95	84	86
蛔虫感染 Infected with Ascaris lumbricoides	10	2	4
鞭虫感染 Infected with Trichuris trichiura	11	1	4
日本血吸虫感染 Infected with Schistosoma japonicum	10	0	1
弓形虫感染 Infected with Toxoplasma gondii	10	0	0
健康者Healthy participants	74	3	4

试条法 Immunochromatographic strip test	ELISA法 Emzyme-linked immunosorbent assay		合计 Total
试条法 Immunochromatographic strip test	阳性Positive	阴性Negative	合计 Total
阳性Positive	84	0	84
阴性Negative	2	9	11
合计 Total	86	9	95