Chilean IPNV isolates: Robustness analysis of PCR detection

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• 作者：Esteban Jorquera^a ; ^d ; Paz Morales^a ; David Tapia^a ; ^c ; Pamela Torres^a ; Yoanna Eissler^a ; Juan C. Espinoza^a ; Pablo Conejeros^b ; ^{pablo.conejeros@uv.cl" class="auth_mail" title="E-mail the corresponding author} ; Juan Kuznar^a
• 关键词：IPNV detection ; Mismatch's Tm analysis ; QPCR
• 刊名：Electronic Journal of Biotechnology
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：20
• 期：Complete
• 页码：28-32
• 全文大小：216 K

文摘

The genomes of several infectious pancreatic necrosis viruses (IPNVs) isolated in Chile were sequenced with a single amplification approach for both segments A and B. The resulting sequences were then used to determine the conservation of the primer-binding regions used in polymerase chain reaction (PCR)-based diagnostic methods proposed in the literature. Thus, the robustness of each technique was studied, particularly the eventual effect of further mutations within the primer-binding sites.

Results

On analysis, most methods currently used to detect Chilean IPNV varieties were deemed adequate. However, the primers were designed to be genogroup specific, implying that most detection methods pose some risk of detecting all strains prevalent in the country, due to the coexistence of genogroups 1 and 5.

Conclusions

Negative results must be interpreted carefully given the high genomic variability of IPNVs. Detection techniques (quantitative reverse transcription (qRT)-PCR) based on degenerate primers can be used to minimize the possibilities of false-negative detections.