Department of Microbiology, Assam University, Silchar, Assam, India.
Division of Animal Health, ICAR-RC for NEH Region, Umiam, Barapani, Meghalaya, India.
Trop Anim Health Prod. 2021 Feb 22;53(1):177. doi: 10.1007/s11250-021-02609-2.
Pig farming performs as an intricate part in the socio-economic situation in the north-eastern region of India. This region contributes 38% (3.95 million) of total pigs in India. In spite of this, the region unables to flourish as an enterprise as per the expectation due to a low productivity rate. Porcine infectious pathogens like porcine cirovirus2 (PCV2) and porcine parvovirus (PPV) have a direct economic impact on pig farming through slow growth rate, abortion, and mortality and ultimately maximize the production cost by increasing the usage of antibiotic or antiviral drugs. The veterinary diagnostic infrastructure is a fundamental aspect of the development of livestock status by rapid and effective detection of pathogens. Quantitative PCR (qPCR) is a precise and fast-track technique used for the routine diagnostic method. Hence, we developed a highly precise and comparatively cost-effective SYBR Green reporter dye-based qPCR assay for parallel identification of PCV2 and PPV. In the present assay, the correlation coefficient (R) value was 0.99, and 10 copies of the gene/μl were the least limit of detection (LOD) concerning both viruses. Melt curve analysis of this study represented PCV2-specific melt curve (Tm) at 81.2 °C and PPV-specific melt curve (Tm) at 73.5 °C. Therefore, the assay easily differentiates the true positive amplicons of PCV2 and PPV through specific Tm values. Among the 50 field samples, 26 (52%) samples were PCV2 positive, 18 (36%) samples PPV positive, and 11 (22%) samples were co-infected of both the viruses. This method is cost-effective, precise, and sensitive to diagnose the concurrent or individual infection of the PCV2 and PPV in the pig. Hence, considering the impact of pig farming in the north-eastern part of the country, the present assay gives an unprecedented achievement in disease diagnosis.
养猪业在印度东北部的社会经济状况中扮演着复杂的角色。该地区贡献了印度总猪数的 38%（395 万头）。尽管如此，由于生产力水平低，该地区未能像预期的那样成为一个企业。猪传染性病原体，如猪圆环病毒 2 型（PCV2）和猪细小病毒（PPV），通过减缓生长速度、流产和死亡，对养猪业造成直接的经济影响，并最终通过增加抗生素或抗病毒药物的使用来最大化生产成本。兽医诊断基础设施是快速有效地检测病原体，从而发展畜牧业的一个基本方面。实时定量 PCR（qPCR）是一种用于常规诊断方法的精确和快速的技术。因此，我们开发了一种高度精确且具有成本效益的基于 SYBR Green 报告染料的 qPCR 检测方法，用于平行鉴定 PCV2 和 PPV。在本研究中，该检测方法的相关系数（R）值为 0.99，两种病毒的最低检测限（LOD）均为 10 个拷贝/μl。该研究的熔解曲线分析表示 PCV2 的特异性熔解曲线（Tm）为 81.2°C，PPV 的特异性熔解曲线（Tm）为 73.5°C。因此，该检测方法通过特定的 Tm 值很容易区分 PCV2 和 PPV 的真实阳性扩增子。在 50 个现场样本中，26 个（52%）样本为 PCV2 阳性，18 个（36%）样本为 PPV 阳性，11 个（22%）样本为两种病毒的混合感染。该方法具有成本效益、精确性和敏感性，可用于诊断 PCV2 和 PPV 的并发或单独感染。因此，考虑到养猪业对印度东北部地区的影响，本研究在疾病诊断方面取得了前所未有的成就。
