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Antigen-Capture Enzyme-Linked Immunosorbent Assay for Specific Detection of Mycoplasma pneumoniae
Published on: February 24, 2023
Development of a Highly Specific and Sensitive Latex Agglutination Test for the Detection of Caprine Mycoplasmosis
P Santhiya1, Surya Sankar1, V Niranjana1
1Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala 680 651 India.
Abstract:
The study was carried out for the cultural and molecular diagnosis of caprine mycoplasmosis in Kerala, India and for the development of an antigen detection latex agglutination test (LAT) for its field diagnosis. A total of 126 samples, comprising 100 nasal swabs and 26 lung tissues, were subjected to isolation and polymerase chain reaction (PCR). The PCR assays included Mycoplasma 16S rRNA genus specific PCR, as well as species and cluster specific PCRs targeting the Mycoides cluster, Mycoides group, Mycoplasma mycoides subsp. capri (Mmc), M. capricolum subsp. capripneumoniae (Mccp), M. capricolum subsp. capricolum (Mcc), M. agalactiae, M. ovipneumoniae, M. conjunctivae, and M. arginini. Additionally, a Mmc strain was characterized by whole genome sequencing (WGS) and the same was used for the development of LAT by sensitizing latex beads with hyperimmune serum raised against Mmc for antigen detection. On Mycoplasma 16S rRNA genus specific PCR, 69 samples (64 nasal swabs and five lung tissue) turned positive. Of these, two lung tissues were positive for Mycoides cluster, Mycoides group, and Mycoplasma mycoides subsp. capri (Mmc) specific PCR. On species specific PCRs, 17 nasal swabs were positive for M. agalactiae, 38 for M. ovipneumoniae, and 8 for M. conjunctivae. The two lung tissue samples, which produced typical colonies on modified Hayflick medium, were confirmed as Mmc using species specific PCR. The representative Mmc strain, Ker.TCR. LT used for developing LAT had a genome size of 1,211,756 bp with 945 protein coding genes and is closely related to a goat derived strain, IVB-X as indicated by core genome phylogenetic analysis. Of the 50 nasal swab samples collected from goats with respiratory tract infections for validation of LAT, 17 were positive by PCR and 14 by LAT. In comparison to PCR, the LAT had a sensitivity of 87.5 percent and specificity of 100 percent. The latex agglutination test is a simple, rapid and reliable screening test under field conditions.
Supplementary Information:
The online version contains supplementary material available at https://doi.org/10.1007/s12088-026-01564-6.
