Piñeiro M, Matas-Quintanilla M, Miralles A, Gutiérrez A M
Acuvet Biotech, C/Bari 25 dpdo, 50197 Zaragoza, Spain.
BioVetMed Research Group, Department of Animal Medicine and Surgery, Veterinary School, CEIR Campus Mare Nostrum (CMN), University of Murcia, Espinardo, 30100 Murcia, Spain.
Res Vet Sci. 2024 Nov;180:105432. doi: 10.1016/j.rvsc.2024.105432. Epub 2024 Oct 5.
The purpose of the study is to develop a sensitive assay for the proper quantification of the acute phase protein Pig-MAP in pig saliva samples. A time-resolved immunofluorometric assay (TR-IFMA) was developed using two pig-MAP-specific monoclonal antibodies. The limit of detection of the assay was 4 ng/mL, enough to measure pig-MAP concentration in saliva. Precision was evaluated for saliva samples of low, medium and high concentration, with inter assay CV of 4-14 % and inter-assay CV of 8-20 %. The assay kept linearity under dilution and a method comparison study performed with serum samples showed good correlation with ELISA. Median Pig-MAP concentration in saliva from healthy animals was 19 ng/mL whereas in pigs with different inflammatory conditions was 11 times higher. In the same animals median pig-MAP serum concentrations were 0.72 mg/mL in the healthy group and 4.61 in the diseased group. The Spearman coefficient of correlation between Pig-MAP concentration in serum and saliva was of 0.72. A correlation was also observed between the salivary concentration of pig-MAP and other two acute phase proteins such as haptoglobin (r = 0.62) or C-reactive protein (r = 0.65). The concentration of Pig-MAP in saliva of pigs with severe respiratory disease decreased significantly from a median value of 128 ng/mL at the time of disease detection to 8 ng/mL after 1 day of antibiotic therapy. Studies performed show that pig-MAP is present in saliva and this specimen may be an alternative to serum for pig-MAP quantification.
本研究的目的是开发一种灵敏的检测方法，用于准确量化猪唾液样本中的急性期蛋白猪-MAP。使用两种猪-MAP特异性单克隆抗体开发了一种时间分辨免疫荧光测定法（TR-IFMA）。该检测方法的检测限为4 ng/mL，足以测量唾液中的猪-MAP浓度。对低、中、高浓度的唾液样本进行了精密度评估，批内变异系数为4%-14%，批间变异系数为8%-20%。该检测方法在稀释条件下保持线性，与血清样本进行的方法比较研究表明与ELISA具有良好的相关性。健康动物唾液中猪-MAP的中位浓度为19 ng/mL，而在患有不同炎症的猪中则高出11倍。在同一动物中，健康组猪-MAP血清中位浓度为0.72 mg/mL，患病组为4.61 mg/mL。血清和唾液中猪-MAP浓度之间的Spearman相关系数为0.72。还观察到猪-MAP唾液浓度与其他两种急性期蛋白如触珠蛋白（r = 0.62）或C反应蛋白（r = 0.65）之间存在相关性。患有严重呼吸道疾病的猪唾液中猪-MAP的浓度在疾病检测时中位数为128 ng/mL，在抗生素治疗1天后降至8 ng/mL，显著降低。进行的研究表明猪-MAP存在于唾液中，该样本可能是用于猪-MAP定量的血清替代物。
