Serodiagnostic Comparison Between Two Methods, ELISA and Surface Plasmon Resonance for the Detection of Antibody Titres of Mycoplasma hyopneumoniae

Abstract

A protein chip based on surface plasmon resonance (SPR) was developed for measuring the Mycoplasma hyopneumoniae antibody titres using a recombinant 30-kDa fragment of P97 adhesin as an antigen. The diagnostic potential of this SPR assay, for detecting the antibody titres to the M. hyopneumoniae 30-kDa protein, was compared with that of conventional ELISA using 70 pig serum samples taken from six pig farms. The SPR assay was found to be highly specific and sensitive. Moreover, there was a strong positive correlation between the SPR and ELISA titres (n = 70, r = 0.898, P < 0.01). Therefore, this recombinant 30-kDa protein can be used as an antigen for serological studies, and the SPR, which is a label-free method, is expected to be a valuable and reproducible tool in the serodiagnosis of M. hyopneumoniae infection.

Development of serodiagnostic surface plasmon resonance imaging assay for the detection of antibodies to porcine circovirus type 2

“…This comparative study showed that although ELISA is quite reliable, it is time-consuming. Recently, many SPR assays have been described for detection of antibodies against pathogens such as porcine circovirus type 2 , classical swine fever virus , Mycoplasma hyopneumoniae (Kim et al, 2006) and Mycoplasma synoviae (Oh et al, 2010). However, the SPR assay can only be applied to 2 to 4 samples simultaneously.…”

Current Issues on Molecular and Immunological Diagnosis of Tuberculosis

“…However, subjective reading of the flow test results is a major drawback. Recently, a fluorescence polarization assay74 and surface plasmon resonance systems75 have been introduced for the detection of antibodies, which can be objectively read. However, the sensitivity and specificity have not been determined in serologic tests for TB.…”

Comparison of Surface Plasmon Resonance Imaging and Enzyme-Linked Immunosorbent Assay for the Detection of Antibodies against Iridovirus in Rock Bream (Oplegnathus Fasciatus)

“…17 SPRI detects the changes in the refractive index within a short distance from the surface of a thin metal film as variations in the light intensity reflected from the back of the film and therefore does not require labeling. 2,3,9 SPRI has been successfully used to screen various bioaffinity interactions using proteins. 8 In this study, the 50-kDa fragment of the major capsid protein (MCP) gene 10 was used to develop a protein chip based on SPRI for detecting antibodies to iridovirus in rock bream sera.…”

“…Recently, many SPR assays have been described for detection of antibodies against pathogens such as porcine circovirus type 2, 2 classical swine fever virus, 3 and Mycoplasma hyopneumoniae. 9 However, the SPR assay can be applied only to 2 to 4 samples simultaneously. Compared with the SPR assay, the newly developed SPRI protein chip with miniaturized micro wells proved to be a rapid (with an assay time of 1 hour), high-throughput, and valuable tool for the serodiagnosis of rock bream iridoviral infection.…”

“…9 Briefly, the MCP gene (GenBank no. AY310918) was amplified by PCR using previously described primers (Table 1).…”