Avian mycoplasmosis, mainly caused by Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) is an economically important disease of the poultry industry. The present study was aimed to develop whole cell based indirect-ELISA (i-ELISA) and DOT blot assay (DOT-ELISA) as rapid, sensitive, specific and economical sero-detection tests for MG and MS. A total of 306 blood samples were collected from birds slaughtered at local meat shops of different districts of Haryana, India to detect MG and MS antibodies. Sonicated antigens prepared from freshly grown culture of MG and MS were used to develop i-ELISA and DOT blot assay. In i-ELISA, 50.32% and 61.76% serum samples were found to be positive for MG and MS antibodies, respectively. However in DOT blot assay, 41.83% and 53.92% serum samples were found positive for MG and MS antibodies, respectively. The relative diagnostic sensitivity and specificity of DOT-ELISA were measured considering i-ELISA as a reference test. The relative diagnostic sensitivity of the DOT blot assay was found to be 69.48% and 82.01%; whereas relative diagnostic specificity was 86.18% and 91.45% for the detection of MG and MS antibodies, respectively. The developed serological assays may be used as rapid and economical diagnostic tools for large scale screening of poultry sera for MG and MS antibodies.

禽支原体病，主要由鸡毒支原体（MG）和滑膜支原体引起(MS) 是家禽业的一种经济上重要的疾病。本研究旨在开发基于全细胞的间接 ELISA (i-ELISA) 和 DOT 印迹试验 (DOT-ELISA) 作为 MG 和 MS 的快速、灵敏、特异和经济的血清检测试验。从印度哈里亚纳邦不同地区的当地肉店屠宰的鸟类中采集了 306 份血液样本，以检测 MG 和 MS 抗体。从新鲜生长的 MG 和 MS 培养物中制备的超声处理抗原用于开发 i-ELISA 和 DOT 印迹分析。在 i-ELISA 中，分别发现 50.32% 和 61.76% 的血清样本对 MG 和 MS 抗体呈阳性。然而，在 DOT 印迹分析中，分别发现 41.83% 和 53.92% 的血清样本对 MG 和 MS 抗体呈阳性。DOT-ELISA 的相对诊断敏感性和特异性是考虑将 i-ELISA 作为参考测试来测量的。DOT印迹法的相对诊断灵敏度分别为69.48%和82.01%；而检测 MG 和 MS 抗体的相对诊断特异性分别为 86.18% 和 91.45%。开发的血清学检测可用作大规模筛查家禽血清中 MG 和 MS 抗体的快速且经济的诊断工具。