Rapid visual detection of tick borne Anaplasma marginale in Indian buffaloes using loop mediated isothermal amplification assay targeting major surface protein 5 gene

利用靶向主要表面蛋白5基因的环介导等温扩增检测法,快速可视化检测印度水牛体内蜱传边缘无形体病。

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Abstract

This study demonstrates the efficacy of loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assays, both targeting the major surface protein 5 (msp5) gene, for the detection of Anaplasma marginale in Indian water buffaloes. The LAMP assay exhibited superior sensitivity, detecting the pathogen in 24 out of 110 samples (21.81%), compared to PCR, which identified 16 out of 110 samples (14.55%), and conventional microscopic examination, which detected only 6 out of 110 samples (5.45%). Relative to PCR, the LAMP assay achieved a diagnostic sensitivity of 93.75% (95% CI 69.77-99.84%) and specificity of 90.43% (95% CI 82.60-95.53%), with a positive predictive value of 62.50% and a negative predictive value of 98.84%. These findings highlight the LAMP assay as a sensitive, specific, and practical diagnostic tool for A. marginale in buffaloes, particularly suitable for field conditions due to its simplicity and visual detection capabilities.

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