Veterinary parasitology, regional studies and reports · 2025
Development and field validation of an indirect ELISA for serosurveillance of Ornithodoros lahorensis exposure.
СтатьяВалидационное исследованиеНазвание и аннотация: Перевод в очереди
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Xu Xiaolin, Yuan Xiangfen, Wang Huiyu, Maimaiti Ailixire, Kong Yufang, Lv Jizhou, Wu Shaoqiang
Аннотация
BACKGROUND: Ornithodoros lahorensis is the primary vector of numerous tick-borne diseases. Effective tick surveillance could facilitate the prevention of tick-borne diseases. Direct surveillance methods for tick bites are impractical, the development of an indirect approach to detect specific antibodies against tick salivary proteins in animal samples would provide a more convenient surveillance and diagnostic tool. Previous studies have confirmed that some salivary gland proteins can be used as detection markers for tick bites.
METHODS: The recombinant BTSP protein of O. lahorensis (rOl-BTSP) was expressed and purified. rOl-BTSP was subjected to sequence alignment analysis, and validated as a serological detection target for O. lahorensi exposure by Western blot analysis. Then, a novel ELISA method for detecting O. lahorensis bites in sheep (S-iELISA) was developed by optimizing critical parameters, including antigen coating concentration, serum dilution. The sensitivity and specificity of the S-iELISA method were evaluated. And the S-iELISA were further applied to test 455 sheep serum samples from Xinjiang and Inner Mongolia in China.
RESULTS: Sequence homology analysis revealed that Ol-BTSP exhibited only 30.5 % amino acid identity with its homologs from both hard ticks and soft ticks. And the serum from mice bitten by O. lahorensis exhibited specific immunoreactivity with rOl-BTSP, while the cross-species antibody tests showed no reactivity, confirming the diagnostic potential of Ol-BTSP for detecting O. lahorensis exposure. The developed S-iELISA exhibited excellent diagnostic accuracy, with a sensitivity of 93.3 % (95 % CI: 70.2-99.7 %) and a specificity of 96.7 % (95 % CI: 82.8-99.9 %) in validation studies. The seroprevalence rate of O. lahorensis exposure in serum samples collected from Xinjiang and Inner Mongolia were 10.45 % and 3.09 %, respectively.
CONCLUSIONS: The S-iELISA method established in this study was a reliable tool for detecting and monitoring O. lahorensis exposure in sheep. And the serological diagnostic method based on rOl-BTSP showed great promise for enhancing the surveillance and management strategies for O. lahorensis-borne diseases.
METHODS: The recombinant BTSP protein of O. lahorensis (rOl-BTSP) was expressed and purified. rOl-BTSP was subjected to sequence alignment analysis, and validated as a serological detection target for O. lahorensi exposure by Western blot analysis. Then, a novel ELISA method for detecting O. lahorensis bites in sheep (S-iELISA) was developed by optimizing critical parameters, including antigen coating concentration, serum dilution. The sensitivity and specificity of the S-iELISA method were evaluated. And the S-iELISA were further applied to test 455 sheep serum samples from Xinjiang and Inner Mongolia in China.
RESULTS: Sequence homology analysis revealed that Ol-BTSP exhibited only 30.5 % amino acid identity with its homologs from both hard ticks and soft ticks. And the serum from mice bitten by O. lahorensis exhibited specific immunoreactivity with rOl-BTSP, while the cross-species antibody tests showed no reactivity, confirming the diagnostic potential of Ol-BTSP for detecting O. lahorensis exposure. The developed S-iELISA exhibited excellent diagnostic accuracy, with a sensitivity of 93.3 % (95 % CI: 70.2-99.7 %) and a specificity of 96.7 % (95 % CI: 82.8-99.9 %) in validation studies. The seroprevalence rate of O. lahorensis exposure in serum samples collected from Xinjiang and Inner Mongolia were 10.45 % and 3.09 %, respectively.
CONCLUSIONS: The S-iELISA method established in this study was a reliable tool for detecting and monitoring O. lahorensis exposure in sheep. And the serological diagnostic method based on rOl-BTSP showed great promise for enhancing the surveillance and management strategies for O. lahorensis-borne diseases.