Key Laboratory of Animal virology of the Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, Gansu, PR China.
BMC Microbiol. 2014 Jan 17;14:10. doi: 10.1186/1471-2180-14-10.
Capripox viruses are economically important pathogens in goat and sheep producing areas of the world, with specific focus on goat pox virus (GTPV), sheep pox virus (SPPV) and the Lumpy Skin Disease virus (LSDV). Clinically, sheep pox and goat pox have the same symptoms and cannot be distinguished serologically. This presents a real need for a rapid, inexpensive, and easy to operate and maintain genotyping tool to facilitate accurate disease diagnosis and surveillance for better management of Capripox outbreaks.
A LAMP method was developed for the specific differential detection of GTPV and SPPV using three sets of LAMP primers designed on the basis of ITR sequences. Reactions were performed at 62°C for either 45 or 60 min, and specificity confirmed by successful differential detection of several GTPV and SPPV isolates. No cross reactivity with Orf virus, foot-and-mouth disease virus (FMDV), A. marginale Lushi isolate, Mycoplasma mycoides subsp. capri, Chlamydophila psittaci, Theileria ovis, T. luwenshuni, T. uilenbergi or Babesia sp was noted. RFLP-PCR analysis of 135 preserved epidemic materials revealed 48 samples infected with goat pox and 87 infected with sheep pox, with LAMP test results showing a positive detection for all samples. When utilizing GTPV and SPPV genomic DNA, the universal LAMP primers (GSPV) and GTPV LAMP primers displayed a 100% detection rate; while the SPPV LAMP detection rate was 98.8%, consistent with the laboratory tested results.
In summary, the three sets of LAMP primers when combined provide an analytically robust method able to fully distinguish between GTPV and SPPV. The presented LAMP method provides a specific, sensitive and rapid diagnostic tool for the distinction of GTPV and SPPV infections, with the potential to be standardized as a detection method for Capripox viruses in endemic areas.
山羊痘病毒和绵羊痘病毒是世界范围内山羊和绵羊养殖区具有经济重要性的病原体，尤其关注山羊痘病毒（GTPV）、绵羊痘病毒（SPPV）和牛结节疹病毒（LSDV）。临床上，绵羊痘和山羊痘的症状相同，无法通过血清学区分。因此，非常需要一种快速、廉价、易于操作和维护的基因分型工具，以促进准确的疾病诊断和监测，从而更好地管理山羊痘爆发。
根据 ITR 序列设计了三套 LAMP 引物，开发了一种用于 GTPV 和 SPPV 特异性差异检测的 LAMP 方法。反应在 62°C 下进行 45 或 60 分钟，通过成功地对几种 GTPV 和 SPPV 分离株进行差异检测来确认特异性。与口疮病毒、口蹄疫病毒（FMDV）、A. marginale Lushi 分离株、Mycoplasma mycoides subsp. capri、鹦鹉热衣原体、绵羊泰勒虫、绵羊泰勒虫、绵羊泰勒虫或巴贝斯虫无交叉反应。对 135 份保存的流行材料进行 RFLP-PCR 分析显示，48 份样本感染山羊痘，87 份样本感染绵羊痘，LAMP 检测结果显示所有样本均为阳性。利用 GTPV 和 SPPV 基因组 DNA 时，通用 LAMP 引物（GSPV）和 GTPV LAMP 引物的检测率为 100%；而 SPPV LAMP 的检测率为 98.8%，与实验室检测结果一致。
综上所述，这三套 LAMP 引物结合使用提供了一种能够完全区分 GTPV 和 SPPV 的分析稳健方法。所提出的 LAMP 方法为区分 GTPV 和 SPPV 感染提供了一种特异性、敏感性和快速的诊断工具，具有在流行地区作为山羊痘病毒检测方法标准化的潜力。
