Suryawanshi Rahul D, Malik Satya Veer Singh, Jayarao Bhushan, Chaudhari Sandeep P, Savage Emily, Vergis Jess, Kurkure Nitin V, Barbuddhe Sukhadeo B, Rawool Deepak B
Division of Veterinary Public Health, ICAR-Indian Veterinary Research Institute, Izatnagar 243 122, India.
Department of Veterinary and Biomedical Sciences, Pennsylvania State University, PA 16801, USA.
J Microbiol Methods. 2017 Jun;137:40-45. doi: 10.1016/j.mimet.2017.04.005. Epub 2017 Apr 12.
The present study for the first time evaluates the serodiagnostic efficacy of two recombinant antigens namely, listeriolysin O (rLLO) and phosphatidyl-inositol phospholipase C (rPI-PLC). Indirect ELISA with the above recombinant antigens was used on samples collected from bovines (n=106), goats (n=138) and pigs (n=92) having either a history of abortion, emaciation and/or apparently healthy animals. Isolation of Listeria was attempted from the blood samples using USDA-FSIS method. On screening of test sera by rLLO-based ELISA, antibodies against anti-listeriolysin O (ALLO) were observed in goats (22.46%), bovines (15.10%) and pigs (16.31%). As advocated, after adsorption of positive serum samples with streptolysin O (SLO), the seropositivity for ALLO was marginally reduced (p>0.05) in goats (21.73%) and bovines (10.38%), whereas, in pigs the reduction (5.43%) was significant (p<0.05). On the contrary, rPI-PLC-based ELISA revealed higher non-specific seropositivity for antilisterial antibodies in goats (45.65%), bovines (31.13%) and pigs (8.69%). Further, on comparing the seropositivity with isolation rate, of the 16 animals that were culturally-positive for L. monocytogenes, 15 showed ALLO positivity in unadsorbed as well as SLO-adsorbed sera by rLLO-based ELISA, however, rPI-PLC-based ELISA could detect seropositivity in only 5 animals. Moreover, rPI-PLC-based ELISA also showed seropositivity in those animals (7/30) that were culturally positive for other Listeria spp. In conclusion, rLLO can serve as a better antigen than rPI-PLC in ELISA for the serodiagnosis of listeriosis in animals; however, prior adsorption of test sera with SLO is required to avoid false positive results.
本研究首次评估了两种重组抗原,即李斯特菌溶血素O(rLLO)和磷脂酰肌醇磷脂酶C(rPI-PLC)的血清学诊断效力。使用上述重组抗原的间接ELISA法检测从有流产、消瘦病史的牛(n=106)、山羊(n=138)和猪(n=92)以及明显健康的动物采集的样本。尝试采用美国农业部食品安全检验局(USDA-FSIS)方法从血样中分离李斯特菌。通过基于rLLO的ELISA法筛查检测血清时,在山羊(22.46%)、牛(15.10%)和猪(16.31%)中观察到抗李斯特菌溶血素O(ALLO)抗体。如所主张的,用链球菌溶血素O(SLO)吸附阳性血清样本后,山羊(21.73%)和牛(10.38%)中ALLO的血清阳性率略有降低(p>0.05),而在猪中降低幅度(5.43%)显著(p<0.05)。相反,基于rPI-PLC的ELISA法显示山羊(45.65%)、牛(31.13%)和猪(8.69%)中抗李斯特菌抗体的非特异性血清阳性率更高。此外,在将血清阳性率与分离率进行比较时,在16头单核细胞增生李斯特菌培养阳性的动物中,15头通过基于rLLO的ELISA法在未吸附以及SLO吸附血清中显示ALLO阳性,然而,基于rPI-PLC的ELISA法仅能检测到5头动物的血清阳性。此外,基于rPI-PLC的ELISA法在那些对其他李斯特菌属培养阳性的动物(7/30)中也显示血清阳性。总之,在ELISA法中,rLLO作为抗原在动物李斯特菌病血清诊断方面比rPI-PLC更好;然而,需要用SLO预先吸附检测血清以避免假阳性结果。