Serological Diagnostic Assays for Detection of Ns1 Antigen, IGM and IGG Antbodies to Dengue Virus
International Journal of Infectious Diseases and Therapy
Volume 2, Issue 1, March 2017, Pages: 9-14
Received: Dec. 18, 2016;
Accepted: Jan. 10, 2017;
Published: Feb. 15, 2017
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Purimitla Usha Rani, Department of Microbiology, GITAM Institute of Medical Sciences and Research, GITAM University, Visakhapatnam, Andhra Pradesh, India
Payala Vijayalakshmi, Department of Microbiology, GITAM Institute of Medical Sciences and Research, GITAM University, Visakhapatnam, Andhra Pradesh, India
Complications in managing the dengue virus infections include the lack of rapid diagnostic procedures and at the same time the symptoms of dengue infection are often confused with those of other diseases. Two commercial rapid serological diagnostic kit methods (Dengue Day 1 test, J Mitra and Co. Pvt. Ltd., New Delhi, ImmunoComb II Dengue IgM/IgGBispot kit (Orgenics Pvt. Ltd., Israel)were evaluated for the detection of NS1 antigen, Immunoglobulin IgG and IgM specific to dengue virus in the serum samples of patients suffered with dengue acute primary infection and secondary infection. The total assay time was 20 min-2hrs. The results of these methods were compared with the gold standard assay methods Dengue IgM-Capture Microplate ELISA and Dengue Indirect IgG ELISA (Pan Bio, Brisbane, Australia). The total assay time was 6-7hrs. Nine serum samples were positive to NS1 antigen and negative to IgG and IgM by Dengue day 1 test. The results of Bispot assay method revealed that, the number of IgG positive samples was 11, IgM positive samples were 31 and both IgG and IgM positive samples were 8. Majority of the positive cases were noticed in the age group 35-68 years and males were more prone to dengue infection while comparing with females. By performing IgM MICROLISA, 34 samples were positive which in turn indicated that, three of them were false negative by the immune comb bispot method giving a sensitivity of 91.17%. Through indirect IgG ELISA, the number of positive samples was 15 and four of the 15 positive samples of IgG were false negative by the immuo comb bispot method giving a sensitivity of 73.33%. The gold standard ELISA methods were more efficient than rapid serological tests and gave an overall sensitivity of 99%. Thus the alternative of an assay that is to be used in the diagnosis of dengue infections depends on factors like laboratory infrastructure, preference and availability of equipment. The allied performance of the Rapid test, followed by confirmation with MAC-ELISA on those samples, ensures both speediness as well as quality of reported results.
Purimitla Usha Rani,
Serological Diagnostic Assays for Detection of Ns1 Antigen, IGM and IGG Antbodies to Dengue Virus, International Journal of Infectious Diseases and Therapy.
Vol. 2, No. 1,
2017, pp. 9-14.
Om Prakash, Rafidah Hanim Shueb. Diagnosis of dengue infection using Conventional and Biosensor based techniques. Viruses. 2015. 7: 5410-5427.
Blacksell SD. Commercial dengue rapid diagnostic tests for point-of-care application: Recent evaluations and future needs. J Biomed. Biotechnol. 2012. 2102.
Guzman MG, Kouri G. Dengue diagnosis, advances and challenges. Int J Infectious diseases. 2004. 8: 69-80.
Natasha Evelyn Anne Murray, Quam MB, Wilder-Smith A. Epidemiology of dengue: past, present and future prospects. Clinical epidemiology. 2013. 5: 299-309.
Pan America Health Organization. Dengue and Dengue hemorrhagic fever in the Americas: Guidelines for prevention and control. 1994. Scientific publication. 548.
Nimmannitya S. Clinical spectrum and management of dengue hemorrhagic fever. Southeast Asian J Trop Med Pub Health. 1987. 20: 325-330.
Martinez E. Dengue hemorragico en criancas. Editorial Jose Marti, La Habana. 1992. 1-180.
Halstead SB. Dengue viruses. In: Gorbach SI, Bartlett JG and Blacklow MR, editors. Infect Dis. Philadelphia: Saunders. 1992. 281-304.
Rothman AL, Ennis FA. Toga/Flaviviruses: Immuno pathology. In: Cunningham MW, Fujinami RS editors. Effects of microbes on the immune system. Philadelphia: Lippincott Williams and Wilkins. 2000. 473-487.
Guzman MG, Kouri G, Valdes L, Bravo J, Vazquez S, Halstead SB. Effect of age outcome of secondary dengue 2 infections. Int J Infect Dis. 2002. 6: 118-124.
Gubler DJ, Rosen L. A simple technique for demonstrating transmission of dengue virus by mosquitoes without the use of vertebrate hosts. Am J Trop Med Hyg. 1977. 26, 533-537.
Carrington LB, Simmons CP. Human to mosquito transmission of dengue viruses. Frontiers in Immunology. 2014. 5: 1-8.
Innis B, Nisalak A, Nimmannitya S, Kusalerdchariya S, Chongswasdi V, Suntayakorn S. An enzyme- linked immune sorbent assay to characterize dengue infections where dengue and Japanese encephalitis co-circulate. Am J Trop Med Hyg. 1989. 40: 418-427.
Nawa M, Takasaki T, Yamada KI, Akatsuka T, Kurane I. Development of Dengue IgM-capture Enzyme-linked Immunosorbent assay with higher sensitivity using monoclonal detection antibody. J Virol Methods. 2001. 92: 65-70.
Laue T, Emmerich P, Schimtz H. Detection of Dengue virus RNA in patients after primary or secondary dengue infection by using the Taqman automated amplification system. J clin Microbiol. 1999. 37: 2543-2547.
Pie-Yun Shu, Huang JH. Current advances in Dengue diagnosis. Clin Diagnostic Lab Immunol. 2004. 11: 642-650.
Tricou V, Vu HT, Quynh NV, Nguyen CV, Tran HT, Farrar J, Wills B, Simmons CP. Comparison of two dengue NS1 rapid tests for sensitivity, Specificity and relationship to Viraemia and antibody responses. BMC Infect Dis. 2010. 10: 142.
Abeyewickreme W, Wickremasinghe AR, Karunatilake, Sommerfeld J, Kroeger A. Community mobilization and household level waste management for dengue vector control in Gampaha district of Sri Lanka; an intervention study. Pathogens and Global Health. 2012. 106 (8): 479-487.
Laferte J, Pelegrino JL, Guzman MG, Gonzalez G, Vazquez S, Hermida C. Rapid diagnosis of dengue virus infection using a novel 10µl IgM antibody capture Ultra MicroElisa assay. Adv Mod Biotech. 1992. 1 (19): 4.
Kuno G, Gomez I, Gubler DJ. An ELISA procedure for the diagnosis of dengue infections. J Virol methods. 1991. 33: 101-113.
Rivetz B, Siman-Tov D, Ambal E, Jaramillo AC, Ben-Zvi A, Tartakovsky B, Falk Fish. New dengue antibody assay with unique differential detection of IgG and IgM antibodies. ClinBiochem. 2009. 42. 180-184.
Wu SJ, Paxton H, Hanson B, Kung CG, Chen TB, Rossi C, Vaughn DW, Murphy G, Hayes C. Comparison of two rapid diagnostic assays for detection of Immunoglobulin IgM antibodies to dengue virus. ClinDiagn Lab Immunol. 2000. 7 (1): 106-110.
Vazquez S, Lemos G, Pupo M et al. Diagnosis of Dengue virus infection by the visual and simple AuBio Dot Immunoglobulin M capture system. ClinDiagn Lab Immunol. 2003. 10 (6): 1074-1077.
Lam SK, Devine PL. Evaluation of capture ELISA and rapid immuno chromatographic testfor the detection of IgG and IgM antibodies produced during dengue infection. ClinDiagnVirol. 1998. 10: 75-81.
Sathish N, Vijayakumar TS, Abraham P, Sridharan G. Dengue fever: its laboratory diagnosis, with special emphasis on IgM detection. Dengue Bulletin. 2003. 27: 116-125.
Mahesh Reddy R, Sahai K, Ajay Malik, Shoba S, Khora A. Comparative analysis of rapid dengue testing for NS1Ag and IgM in acute dengue infection. Int J Current Microbiol Appl Sci. 2016. 5 (10): 931-937.
Narayan R, Raja S, Kumar S, Sambasivam M, Jagadersan R, Arunagiri K, Krishnasamy K, Palani G. A novel Indirect ELISA for diagnosis of dengue fever. Ind J Med Res. 2016. 144 (1): 128-133.
Manthalkar PS, Peerapur BV. Sero diagnosis of dengue virus infection using ELISA in patients with suspected dengue infection. J Evolution Med Dent Sci. 2015. 4 (62): 10824-10828.
Kong YY, Thayan R, Chong HT, Tan CT, Devi S. Rapid detection and serotyping of dengue virus by Multiplex RT-PCR and Real-Time SYBR Green RT-PCR. Sing Med J. 2007. 48: 662-668.