APPLICATION OF EXPRESSED RECOMBINANT OUTER MEMBRANE PROTEIN (OMPA PROTEIN) FROM BURKHOLDERIA THAILANDENSIS E264 IN DOT BLOT ELISA

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Kanwitchayut Sroiphetcharat
Sumet Wajanarogana
Kanyanan Kritsiriwuthinan
Patamaporn Sukplang
Acharawan Thongmee

Abstract

The recombinant OmpA protein from Burkholderia thailandensis, the closely linked species to Burkholderia pseudomallei, was expressed in E. coli BL21 (DE3).  The purified OmpA protein was assessed as an antigen that specifically reacts to melioidosis antibodies by dot blot ELISA.  In this study implementations were practiced with 5 bacterial-culture-confirmed melioidosis sera and 3 healthy donor sera.  At the serum dilutions of 1:800, 1:1,600 and 1:3,200, the sensitivities of 100%, 100%, 60% and specificities of 0%, 0%, 100% were shown, respectively.  Therefore, it can be concluded that the B. thailandensis OmpA protein antigen developed may possibly be used for a serological diagnosis of melioidosis by dot blot ELISA at the cut-off titer of 3,200 with sensitivity and specificity of 60% and 100%.

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How to Cite
1.
Sroiphetcharat K, Wajanarogana S, Kritsiriwuthinan K, Sukplang P, Thongmee A. APPLICATION OF EXPRESSED RECOMBINANT OUTER MEMBRANE PROTEIN (OMPA PROTEIN) FROM BURKHOLDERIA THAILANDENSIS E264 IN DOT BLOT ELISA. Interprof J Health Sci [Internet]. 2023 Oct. 5 [cited 2024 Dec. 22];15(2):54-62. Available from: https://li05.tci-thaijo.org/index.php/IJHS/article/view/174
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Research Articles

References

Allwood, E.M., Logue, C.A., Hafner, G.J., Ketheesan, N., Norton, R.E., Peak, I.R., and Beacham, I.R. 2008. Evaluation of recombinant antigens for diagnosis of melioidosis. FEMS Immunol. Med. Microbiol. 54: 144-153.

Anandan, S., Augustine, A., Mathai, E., and Jesudason, M.V. 2010. Evaluation of IgM ELISA using a sonicate and a lipopolysaccharide antigen for the serodiagnosis of melioidosis. Indian J. Med. Microbiol. 28: 158-161.

Anuntagool, N., Naigowit, P., Petkanchanapong, V., Aramsri, P., Panichakul, T., and Sirisinha, S. 2000. Monoclonal antibody-based rapid identification of Burkholderia pseudomallei in blood culture fluid from patients with community-acquired septicaemia. J. Med. Microbiol. 49: 1075-1078.

Cheng, A.C., O’Brien, M., Freeman, K., Lum, G., and Currie, B.J. 2006. Indirect hemagglutination assay in patients with melioidosis in northern Australia. Am. J. Trop. Med. Hyg. 74: 330-334.

Confer, A.W., and Ayalew, S. 2013. The OmpA family of proteins: Roles in bacterial pathogenesis and immunity. Vet. Microbiol. 163: 207-222.

Desakorn, V., Smith, M.D., Wuthiekanun, V., Dance, D.A., Aucken, H., Suntharasamai, P., Rajchanuwong, A., and White, N.J. 1994. Detection of Pseudomonas pseudomallei antigen in urine for the diagnosis of melioidosis. Am. J. Trop. Med. Hyg. 51: 627-633.

Dharakul, T., Songsirivilai, S., Viriyachitra, S., Luangwedchakarn, V., Tassaneetritap, B., and Chaowagul, W. 1996. Detection of Burkholderia pseudomallei DNA in Patients with Septicemic Melioidosis. J. Clin. Microbiol. 34: 609-614.

Hara, Y., Moham, R., Nathan, S. 2009. Immunogenic Burkholderia pseudomallei outer membrane proteins as potential candidate vaccine targets. PloS One 4(8):e6496.

Limmathurotsakul, D., Jamsen, K., Arayawichanont, A., Simpson, J.A., White, L.J., Lee, S.J., Wuthiekanun, V., Chantratita, N., Cheng, A., Day, N.P., Verzilli, C., and Peacock, S.J. 2010. Defining the true sensitivity of culture for the diagnosis of melioidosis using Bayesian latent class models. PLoS One. 5: e12485.

Parikn, R., Mathai, A., Parikh, S., Chandra Sekhar, G. and Thomas, R. 2008. Understanding and using sensitivity, specificity and predictive values. Indian Journal of Ophthalmplogy, 56(1): 45-50.

Parthasarathy, N., Saksena, R., Kováč, P., Deshazer, D., and Peacock, S. J. 2008. Application of carbohydrate microarray technology for the detection of Burkholderia pseudomallei, Bacillus anthracis and Francisella tularensis antibodies. Carbohydr Res., 343: 2783–2788.

Wajanarogana, S., Nimnuch, P., Thongmee, A., and Kritsiriwuthinan, K. 2013. Potential of recombinant flagellin fragment from Burkholderia thailandensis as an antigen for melioidosis antibody detection by indirect ELISA. Mol. Cell. Probes. 27: 98-102.

Wajanarogana, S., and Kritsiriwuthinan, K. 2016. Efficacy of indirect ELISA for serodiagnosis of melioidosis using immunodominant antigens from non‑pathogenic Burkholderia thailandensis. SpringerPlus, 5 (1814): 1-8.

Wuthiekanun, V., Chierakul, W., Langa, S., Chaowagul, W., Panpitpat, C., Saipan, P., Thoujaikong, T., Day, N.P., and Peacock, S.J. 2006. Development of antibodies to Burkholderia pseudomallei during childhood in melioidosis-endemic northeast Thailand. Am. J. Trop. Med. Hyg. 74: 1074 –1075.

Wuthiekanun, V., Desakorn, V., Wongsuvan, G., Amornchai, P., Cheng, A.C., Maharjan, B., Limmathurotsakul, D., Chierakul, W., White, N.J., Day, N.P.J., and Peacock, S.J. 2005. Rapid Immunofluorescence Microscopy for Diagnosis of Melioidosis. Clin. Diagn. Lab. Immunol. 12: 555–556.