A Novel Nano Magnetic Beads Dot ELISA Immunoassay and Its Application on the Detection of Giardia lamblia Coproantigen
,
Abstract
Background: The traditional basis of diagnosis is identification of Giardia lamblia trophozoites or cysts in the stool of infected patients. Recently the advent of more objective techniques like antigen detection methods has led to an increase in their use versus those that rely on subjective microscopic examination of fecal specimens for Giardia cysts may facilitate diagnosis of G. lamblia in stool specimens.
Methods: This cross-sectional study was carried out from Oct 2015 to Feb 2016 on patients admitted to Benha University Hospitals (Benha, Egypt) and outpatients of Theodor Bilharz Research Institute (TBRI) (Giza, Egypt). Purified G. lamblia cysts antigen was prepared by two-phase sucrose gradient technique. Polyclonal antibody against purified G. lamblia cysts antigen was prepared and labeled with horseradish peroxidase and Nano Magnetic Beads (NMB) to be used as detecting antibody. A total of 72 stool samples, 32 samples positive for giardiasis, 20 samples positive for other parasitic infections in addition to 20 negative samples were examined using dot ELISA and NMB dot-ELISA.
Results: The sensitivity of the traditional dot-ELISA was 81.3 % and it increased by using the NMB-dot-ELISA to be 96.9% in stool samples. Specificity of both techniques was 97.5%.
Conclusion: Diagnosis of G. lamblia by NMB-Dot-ELISA technique is sensitive, specific, rapid and easy to perform and interpret. In this study, using the nano-magnetic beads increased the sensitivity of the applied technique.
Feng Y, Xiao L. Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clinical Microbiology Reviews. 2011 Jan 1;24(1):110-40.
Barry MA, Weatherhead JE, Hotez PJ, Woc-Colburn L. Childhood parasitic infections endemic to the United States. Pediatric Clinics of North America. 2013 Apr 30;60(2):471-85.
Coelho CH, Costa AO, Silva AC, Pucci MM, Serufo AV, Busatti HG, Durigan M, Perales J, Chapeaurouge A, e Silva DA, Gomes MA. Genotyping and Descriptive Proteomics of a Potential Zoonotic Canine Strain of Giardia duodenalis, Infective to Mice. PloS one. 2016 Oct 19;11(10):e0164946.
Núñez FA, Hernández M, Finlay CM. Longitudinal study of giardiasis in three day care centres of Havana City. Acta tropica. 1999 Oct 15;73(3):237-42.
Pappas MG. Recent applications of the Dot-ELISA in immunoparasitology. Veterinary parasitology. 1988 Sep 1;29(2-3):105-29.
Venkataramana M, Rashmi R, Uppalapati SR, Chandranayaka S, Balakrishna K, Radhika M, Gupta VK, Batra HV. Development of sandwich dot-ELISA for specific detection of Ochratoxin A and its application on to contaminated cereal grains originating from India. Frontiers in microbiology. 2015;6.
Arjmand J, Esmaeilnejad B, Jalali MH, Ghorbanpoor M, Froushani SM. Designing and evaluation of Dot-ELISA for diagnosis of Fasciola infection in cattle. InVeterinary research forum: an international quarterly journal 2014 (Vol. 5, No. 2, p. 141). Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
Moharm IM, El Enain G, Bahaa El Deen W, Mahgoub A, Hendawy M, Aly I. Evaluation of Nano-Gold Beads Based-ELISA for Detection of Giardia lamblia Antigen in Stool Samples of Infected Patients. Middle-East Journal of Scientific Research. 2014;21(12):2264-73.
Tiwari PM, Vig K, Dennis VA, Singh SR. Functionalized gold nanoparticles and their biomedical applications. Nanomaterials. 2011 Jun 14;1(1):31-63.
El-Ansary A, Al-Daihan S. On the toxicity of therapeutically used nanoparticles: an overview. Journal of Toxicology. 2009 Jan 25;2009.
Blagg W, Schloegel EL, Mansour NS, Khalaf GI. A new concentration technic for the demonstration of protozoa and helminth eggs in feces. American Journal of Tropical Medicine and Hygiene. 1955 Jan;4(1):23-8.
Mezo M, González-Warleta M, Carro C, Ubeira FM. An ultrasensitive capture ELISA for detection of Fasciola hepatica coproantigens in sheep and cattle using a new monoclonal antibody (MM3). Journal of Parasitology. 2004 Aug;90(4):845-52.
Guobadia EE, Fabemi BO. The isolation of Fasciola gigantica-specific antigens and their use in the serodiagnosis of fasciolosis in sheep by the detection of circulating antigens. Veterinary parasitology. 1997 Feb 28;68(3):269-82.
Fagbemi BO, Obarisiagbon IO, Mbuh JV. Detection of circulating antigen in sera of Fasciola gigantica-infected cattle with antibodies reactive with a Fasciola-specific 88-kDa antigen. Veterinary parasitology. 1995 Jun 30;58(3):235-46.
Tijssen P, Kurstak E. Highly efficient and simple methods for the preparation of peroxidase and active peroxidase-antibody conjugates for enzyme immunoassays. Analytical biochemistry. 1984 Feb 1;136(2):451-7.
Mu X, Tong Z, Huang Q, Liu B, Liu Z, Hao L, Zhang J, Gao C, Wang F. Nano-magnetic immunosensor based on staphylococcus protein a and the amplification effect of HRP-conjugated phage antibody. Sensors. 2015 Feb 9;15(2):3896-910.
Engvall E, Perlmann P. Enzyme-linked immunosorbent assay (ELISA) quantitative assay of immunoglobulin G. Immunochemistry. 1971 Sep 1;8(9):871-4.
Rokni MB, Samani A, Massoud J, Nasr MB. Evaluation of Dot-ELISA method using excretory-secretary antigens of Fasciola hepatica in laboratory diagnosis of human fasciolosis. Iranian Journal of Parasitology. 2006;1(1):26-30.
Aly I, Metwalley KM, Nagy F, EL-Mohandes M. Evaluation of Conventional Parasitological Assays, Antigen Detection and Rapid Kit for Diagnosis of Human Giardiasis.
Aly I, Zalat R, El Aswad BW, Moharm IM, Masoud BM, Diab T. Novel nanomagnetic beads based–latex agglutination assay for rapid diagnosis of human schistosomiasis haematobium. Int J Med Health Biomed Pharm Eng. 2013;7(12):642-7.
Catty D, Raykundalia C. ELISA and related enzyme immunoassays. In: Catty D. (Eds.). Antibodies, A practical approach. Vol. II. London, UK: Oxford University, IRL Press 1989; 97-152.
Roxström-Lindquist K, Palm D, Reiner D, Ringqvist E, Svärd SG. Giardia immunity–an update. Trends in parasitology. 2006 Jan 31;22(1):26-31.
Geurden T, Vercruysse J, Claerebout E. Is Giardia a significant pathogen in production animals?. Experimental parasitology. 2010 Jan 31;124(1):98-106.
Jelinek T, Neifer S. Detection of Giardia lamblia stool samples: a comparison of two enzyme-linked immunosorbent assays. F1000Research. 2013 Feb 11;2.
Selim MA, Taha AA, El-Aal NF, Farag TI, Yousef AM. DETECTION OF GIARDIA INTESTINALIS COPROANTIGENS IN DIARRHEIC SAMPLES BY IMMUNOCHROMATOGRAPHIC AND ELISATECHNIQUES. Journal of the Egyptian Society of Parasitology. 2015 Aug;45(2):273-83.
Al Saeed AT, Issa SH. Detection of Giardia lamblia antigen in stool specimens using enzyme-linked immunosorbent assay. Eastern Mediterranean Health Journal. 2010; 362-364.
Sadaka HA, Gaafar MR, Mady RF, Hezema NN. Evaluation of ImmunoCard STAT test and ELISA versus light microscopy in diagnosis of giardiasis and cryptosporidiosis. Parasitology research. 2015 Aug 1;114(8):2853-63.
Singhal S, Mittal V, Khare V, Singh YI. Comparative analysis of enzyme-linked immunosorbent assay and direct microscopy for the diagnosis of Giardia intestinalis in fecal samples. Indian Journal of Pathology and Microbiology. 2015 Jan 1;58(1):69.
Al FD, Kuştimur S, Özekinci T, Balaban N, İlhan MN. The use of enzyme linked immunosorbent assay (ELISA) and direct fluorescent antibody (DFA) methods for diagnosis of Giardia intestinalis. Turkiye Parazitol Derg. 2006;30(4):275-8.
Maraha B, Buiting AG. Evaluation of four enzyme immunoassays for the detection of Giardia lamblia antigen in stool specimens. European Journal of Clinical Microbiology and Infectious Diseases. 2000 Jul 1;19(6):485-7.
Fedorko DP, Williams EC, Nelson NA, Calhoun LB, Yan SS. Performance of three enzyme immunoassays and two direct fluorescence assays for detection of Giardia lamblia in stool specimens preserved in ECOFIX. Journal of clinical microbiology. 2000 Jul 1;38(7):2781-3.
Duguet E, Vasseur S, Mornet S, Devoisselle JM. Magnetic nanoparticles and their applications in medicine.
Gu H, Xu K, Xu C, Xu B. Biofunctional magnetic nanoparticles for protein separation and pathogen detection. Chemical Communications. 2006(9):941-9.
El-Ansary A, Faddah LM. Nanoparticles as biochemical sensors. Nanotechnology, science and applications. 2010 Sep 23;3(1):65-76.
| Files | ||
| Issue | Vol 13 No 4 (2018) | |
| Section | Original Article(s) | |
| Keywords | ||
| Giardia lamblia Dot ELISA Nano technology | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
