Diagnostic utility of intranodular saline lavage in TaqMan probe-based real-time PCR diagnosis of lumpy skin disease

Ajay Pratap Singh , Nidhi Dangi , Prachi Singh , Vinod Kumar Singh , Shyama N Prabhu , Neeraj Kumar Gangwar , Sanchay Kumar Biswas
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Abstract

Lumpy skin disease (LSD), caused by the poxvirus lumpy skin disease virus has entered India, causing serious economic consequences for the Indian dairy sector. To evaluate the diagnostic value of intranodular saline lavage in the identification of LSD-infected animals, skin nodule tissue biopsy, intranodular saline lavage, whole blood, and nasal swab specimens were collected simultaneously from LSD-symptomatic animals. TaqMan probe-based real-time PCR was performed targeting the EEV glycoprotein gene (LSDV126) and the results were compared. qPCR analysis could detect LSDV in all the skin nodule tissue biopsy and intranodular saline lavage. The mean cycle threshold (Cq) values of skin nodule tissue biopsy samples (23.94 ± 2.97) were also comparable with intranodular saline lavage (25.23 ± 2.41) samples. Whole blood and nasal swabs have an extremely poor detection rate for PCR-based viral detection.Our findings demonstrate that intranodular saline lavage is an excellent alternative specimen for the detection of LSDV. Taking into consideration, the simplicity of specimen collection, intranodular saline lavage could be adopted in field conditions for accurate LSDV surveillance testing.

基于 TaqMan 探针的实时 PCR 诊断结节性皮肤病时结节内生理盐水灌洗的诊断作用
由痘病毒引起的疙瘩性皮肤病(LSD)已进入印度,给印度乳制品部门造成严重的经济后果。为了评价生理盐水灌洗对lsd感染动物的诊断价值,我们同时采集lsd症状动物的皮肤结节组织活检、生理盐水灌洗、全血和鼻拭子标本。采用TaqMan探针对EEV糖蛋白基因(LSDV126)进行实时PCR检测,并对结果进行比较。qPCR分析可在所有皮肤结节组织活检和结节内生理盐水灌洗中检测到LSDV。皮肤结节组织活检样本的平均周期阈值(Cq)为23.94±2.97,与结节内生理盐水灌洗样本(25.23±2.41)相当。全血和鼻拭子对pcr病毒检测的检出率极低。我们的研究结果表明,结节内盐水灌洗是检测LSDV的一个很好的替代标本。考虑到标本采集简单,可在野外条件下采用生理盐水灌洗进行准确的LSDV监测检测。
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