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基于重组溶血素O(LLO)和磷脂酰肌醇特异性磷脂酶C(PI-PLC)的酶联免疫吸附测定法(ELISA)在动物李斯特菌病检测中的比较诊断效能

Comparative diagnostic efficacy of recombinant LLO and PI-PLC-based ELISAs for detection of listeriosis in animals.

作者信息

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

机构信息

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.

出版信息

J Microbiol Methods. 2017 Jun;137:40-45. doi: 10.1016/j.mimet.2017.04.005. Epub 2017 Apr 12.

Abstract

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.

摘要

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。

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