全部 标题 作者
关键词 摘要

PLOS ONE  2014 

Preparation of Colloidal Gold Immunochromatographic Strip for Detection of Paragonimiasis skrjabini

DOI: 10.1371/journal.pone.0092034

Full-Text   Cite this paper   Add to My Lib


Background Paragonimiasis is a food-borne trematodiasis, a serious public health issue and a neglected tropical disease. Paragonimus skrjabini is a unique species found in China. Unlike paragonimiasis westermani, it is nearly impossible to make a definitive diagnosis for paragonimiasis skrjabini by finding eggs in sputum or feces. Immunodiagnosis is the best choice to detect paragonimiasis skrjabini. There is an urgent need to develop a novel, rapid and simple immunoassay for large-scale screening patients in endemic areas. Methodology/Principal Findings To develop a rapid, simple immunodiagnostic assay for paragonimiasis, rabbit anti-human IgG was conjugated to colloidal gold particles and used to detect antibodies in the sera of paragonimiasis patients. The synthesis and identification of colloidal gold particles and antibody-colloidal gold conjugates were performed. The size of colloidal gold particles was examined using a transmission electron microscope (TEM). The average diameter of colloidal gold particles was 17.46 nm with a range of 14.32–21.80 nm according to the TEM images. The formation of antibody-colloidal gold conjugates was monitored by UV/Vis spectroscopy. Excretory-secretory (ES) antigen of Paragonimus skrjabini was coated on nitrocellulose membrane as the capture line. Recombinant Staphylococcus protein A was used to prepare the control line. This rapid gold immunochromatographic strip was assembled in regular sequence through different accessories sticked on PVC board. The relative sensitivity and specificity of the strip was 94.4% (51/54) and 94.1% (32/34) respectively using ELISA as the standard method. Its stability and reproducibility were quite excellent after storage of the strip at 4°C for 6 months. Conclusions/Significance Immunochromatographic strip prepared in this study can be used in a rapid one-step immunochromatographic assay, which is instantaneous and convenient.


[1]  Keiser J, Utzinger J (2005) Emerging foodborne trematodiasis. Emerg Infect Dis 11: 1507–1514. doi: 10.3201/eid1110.050614
[2]  Liu Q, Wei F, Liu WS, Yang ST, Zhang XC (2008) Paragonimiasis: an important food-borne zoonosis in China. Trends in Parasitology 24: 318–323. doi: 10.1016/j.pt.2008.03.014
[3]  Zhang ZH (2005) Paragonimiasis. Diseases of Natural Focus Chinese Science Press: 1085–1098.
[4]  Zhang XL, Wang Y, Wang GX, Chen WB, He X, et al. (2012) Distribution and clinical features of Paragonimiasis skrjabini in Three Gorges Reservoir Region. Parasitol Int 61: 645–649. doi: 10.1016/j.parint.2012.06.007
[5]  Chai JY (2013) Paragonimiasis. Handb Clin Neurol 114: 283–296.
[6]  Nkouawa A, Sako Y, Moyou-Somo R, Ito A (2011) Serological and molecular tools to detect neurologic parasitic zoonoses in rural Cameroon. Southeast Asian J Trop Med Public Health 42: 1365–1374.
[7]  Kim YJ, Lee SM, Choi GE, Hwang SH, Kim HH, et al. (2010) Performance of an enzyme-linked immunosorbent assay for detection of Clonorchis sinensis infestation in high- and low-risk groups. J Clin Microbiol 48: 2365–2367. doi: 10.1128/jcm.02506-09
[8]  Zhu YC (2005) Immunodiagnosis and its role in schistosomiasis control in China: a review. Acta Trop 96: 130–136. doi: 10.1016/j.actatropica.2005.07.007
[9]  Rouf MA, Rahman ME, Islam MN, Islam MN, Ferdous NN, et al. (2009) Sensitivity, specificity and predictive values of immunochromatographic strip test in diagnosis of childhood kala-azar. Mymensingh Med J 18: S1–5.
[10]  Zhang GP, Guo JQ, Wang XN, Yang JX, Yang YY, et al. (2006) Development and evaluation of an immunochromatographic strip for trichinellosis detection. Vet Parasitol 137: 286–293. doi: 10.1016/j.vetpar.2006.01.026
[11]  Paek SH, Lee SH, Cho JH, Kim YS (2000) Development of rapid one-step immunochromatographic assay. Methods 22: 53–60. doi: 10.1006/meth.2000.1036
[12]  Frens G (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nature Physical Science 241: 20–22. doi: 10.1038/physci241020a0
[13]  Perfetti M, Pineider F, Poggini L, Otero E, Mannini M, et al.. (2013) Grafting Single Molecule Magnets on Gold Nanoparticles. Small.
[14]  Philip D (2008) Synthesis and spectroscopic characterization of gold nanoparticles. Spectrochim Acta A Mol Biomol Spectrosc 71: 80–85.
[15]  Devi KR, Narain K, Bhattacharya S, Negmu K, Agatsuma T, et al. (2007) Pleuropulmonary paragonimiasis due to Paragonimus heterotremus: molecular diagnosis, prevalence of infection and clinicoradiological features in an endemic area of northeastern India. Trans R Soc Trop Med Hyg 101: 786–792. doi: 10.1016/j.trstmh.2007.02.028
[16]  Odermatt P, Habe S, Manichanh S, Tran DS, Duong V, et al. (2007) Paragonimiasis and its intermediate hosts in a transmission focus in Lao People's Democratic Republic. Acta Trop 103: 108–115. doi: 10.1016/j.actatropica.2007.05.015
[17]  Doanh PN, Shinohara A, Horii Y, Habe S, Nawa Y (2009) Discovery of Paragonimus westermani in Vietnam and its molecular phylogenetic status in P. westermani complex. Parasitol Res 104: 1149–1155. doi: 10.1007/s00436-008-1302-z
[18]  Nkouawa A, Sako Y, Itoh S, Kouojip-Mabou A, Nganou CN, et al. (2010) Serological studies of neurologic helminthic infections in rural areas of southwest cameroon: toxocariasis, cysticercosis and paragonimiasis. PLoS Negl Trop Dis 4: e732. doi: 10.1371/journal.pntd.0000732
[19]  Maltha J, Gillet P, Jacobs J (2013) Malaria rapid diagnostic tests in endemic settings. Clin Microbiol Infect 19: 399–407. doi: 10.1111/1469-0691.12151
[20]  Yan J, Li N, Wei X, Li P, Zhao Z, et al. (2013) Performance of two rapid diagnostic tests for malaria diagnosis at the China-Myanmar border area. Malar J 12: 73. doi: 10.1186/1475-2875-12-73
[21]  Sheele JM, Kihara JH, Baddorf S, Byrne J, Ravi B (2013) Evaluation of a novel rapid diagnostic test for Schistosoma haematobium based on the detection of human immunoglobulins bound to filtered Schistosoma haematobium eggs. Trop Med Int Health 18: 477–484. doi: 10.1111/tmi.12063
[22]  Azzazy HM, Mansour MM (2009) In vitro diagnostic prospects of nanoparticles. Clin Chim Acta 403: 1–8. doi: 10.1016/j.cca.2009.01.016
[23]  Thobhani S, Attree S, Boyd R, Kumarswami N, Noble J, et al. (2010) Bioconjugation and characterisation of gold colloid-labelled proteins. J Immunol Methods 356: 60–69. doi: 10.1016/j.jim.2010.02.007
[24]  Lou S, Ye JY, Li KQ, Wu A (2012) A gold nanoparticle-based immunochromatographic assay: the influence of nanoparticulate size. Analyst 137: 1174–1181. doi: 10.1039/c2an15844b
[25]  Jurgens L, Nichtl A, Werner U (1999) Electron density imaging of protein films on gold-particle surfaces with transmission electron microscopy. Cytometry 37: 87–92. doi: 10.1002/(sici)1097-0320(19991001)37:2<87::aid-cyto1>3.3.co;2-t
[26]  Sheng XZ, Song JL, Zhan WB (2012) Development of a colloidal gold immunochromatographic test strip for detection of lymphocystis disease virus in fish. J Appl Microbiol 113: 737–744. doi: 10.1111/j.1365-2672.2012.05389.x
[27]  Qian X, Peng XH, Ansari DO, Yin-Goen Q, Chen GZ, et al. (2008) In vivo tumor targeting and spectroscopic detection with surface-enhanced Raman nanoparticle tags. Nat Biotechnol 26: 83–90. doi: 10.1038/nbt1377
[28]  Hewitson JP, Grainger JR, Maizels RM (2009) Helminth immunoregulation: the role of parasite secreted proteins in modulating host immunity. Mol Biochem Parasitol 167: 1–11. doi: 10.1016/j.molbiopara.2009.04.008
[29]  Yang SH, Park JO, Lee JH, Jeon BH, Kim WS, et al. (2004) Cloning and characterization of a new cysteine proteinase secreted by Paragonimus westermani adult worms. Am J Trop Med Hyg 71: 87–92.
[30]  Posthuma-Trumpie GA, Korf J, van Amerongen A (2009) Lateral flow (immuno)assay: its strengths, weaknesses, opportunities and threats. A literature survey. Anal Bioanal Chem 393: 569–582. doi: 10.1007/s00216-008-2287-2


comments powered by Disqus