Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an, China.
Parasite Immunol. 2014 Feb;36(2):53-9. doi: 10.1111/pim.12076.
The mite Psoroptes cuniculi is globally widespread and has a serious impact on commercial rabbit breeding. In China, diagnosis of P. cuniculi is currently based on conventional clinical methods that entail numerous disadvantages, including their failure to diagnose subclinical infections. Hence, alternative measures are required, and dot-ELISA is one of the most promising strategies. We cloned and expressed the recombinant P. cuniculi troponin C gene for use as a basis for novel dot-ELISA assay to detect P. cuniculi infections in rabbits. This amplified sequence encoded a 153 amino acid protein of 17·6 kDa and theoretical pI 4·18 without signal peptide. The recombinant troponin C of P. cuniculi is an outer membrane protein and may also be a new P. cuniculi allergen. Results of dot-ELISA test showed that this novel assay had more than 90% sensitivity but low specificity in distinguishing infections with P. cuniculi or Sarcoptic scabiei, despite very high agreement between observers (97-99%; κ values ranged from 0·95 to 0·98 for inter- and intra-observer variability test). This study showed that this novel method, at present, lacks diagnostic utility. Therefore, although simple serological assays such as dot-ELISA show great promise as diagnostic tools, we suggest that troponin C is not a suitable diagnostic antigen candidate.
兔疥螨是一种广泛分布的螨,对商业养兔业有严重影响。目前在中国,兔疥螨的诊断主要基于传统的临床方法,但存在许多缺点,包括无法诊断亚临床感染。因此,需要替代措施,点 ELISA 就是其中最有前途的策略之一。我们克隆并表达了重组兔疥螨肌钙蛋白 C 基因,用于建立新的点 ELISA 检测方法,以检测兔疥螨感染。该扩增序列编码了一个编码 153 个氨基酸的 17.6 kDa 蛋白,理论等电点为 4.18,无信号肽。重组兔疥螨肌钙蛋白 C 是一种外膜蛋白,也可能是一种新的兔疥螨过敏原。点 ELISA 检测结果表明,该新型检测方法对兔疥螨或疥螨感染的敏感性超过 90%,但特异性较低,尽管观察者之间的一致性非常高(观察者间和观察者内变异试验的κ值范围为 0.95 至 0.98)。本研究表明,该方法目前缺乏诊断效用。因此,尽管简单的血清学检测方法如点 ELISA 显示出作为诊断工具的巨大潜力,但我们认为肌钙蛋白 C 不是一种合适的诊断抗原候选物。