Modification and optimization of a microtiter plate (in-house plate coating) for immunoglobulin M (IgM) measles detection by indirect enzyme-linked immunosorbent assay (ELISA)

Abstract

Cases of measles infection in Indonesia are still widespread. Clinical measles infections can only be confirmed at the National Laboratory. The routinely used commercial testing kits are limited. This paper reports the development of a specific in-house plate coating with indirect ELISA. This method involves modifying and optimizing a microtiter plate with measles virus culture. Viral measles culture was obtained by growing it on the cell culture vero/hSLAM. The plate coating was optimized using a culture of measles virus with MO/2007/Jakarta and J/2010/Riau isolates in dilution of 1:1–1:2.048. The virus was inactivated at the time of coating. The indirect ELISA examination for the in-house plate coating was optimized for conjugate concentrations of 1:10, 1:25 and 1:50. The coating showed optimum results for detecting IgM measles at 1:16 dilution with MO/38/V/07 isolates in inactivation and examination of indirect ELISA using a 1:25 conjugate concentration.