Scholarlink

<p>Fruit fly infestation poses a major threat to cucurbit production worldwide, with the melon fruit fly (<i>Zeugodacus cucurbitae</i>) being one of the most severe due to its widespread distribution and significant yield losses. Accurate identification of <i>Z. cucurbitae</i> remains challenging due to overlapping adult morphological characters within cryptic species complexes and difficulties in identifying immature stages. To address this, we developed a species-specific primer (SSP) assay targeting the mitochondrial cytochrome c oxidase subunit I (<i>COI</i>) gene for precise identification of <i>Z. cucurbitae</i>. The SSP was subjected to various tests, including cross-specificity, sensitivity densitometric, and real-time PCR assay. The SSP (ZCRP1F/ZCRP1R) exhibited 100% accuracy across specimens from diverse geographic regions in India and demonstrated high specificity, with no cross-amplification among other economically important fruit fly species. Sensitivity tests confirmed detection limits as low as 10&#xa0;pg/µl in standard PCR and 1&#xa0;pg/µl in real-time PCR, yielding a 294&#xa0;bp amplicon. Furthermore, compatibility with the Real Gene Tissue-Direct PCR kit enabled direct amplification from insect tissues, eliminating the need for DNA extraction. This streamlined approach enhances diagnostic efficiency, reducing processing time. The developed assay has significant potential for applications in quarantine inspections, border security, and for prevention of entry, contributing to improved pest management and biosecurity measures.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Real-time PCR assay for the rapid and precise detection of the melon fly, Zeugodacus cucurbitae (Coquillett) (Diptera: Tephritidae)

  • 期刊论文

  • 发表日期：

    2025年11月6日

查看全文
  • Varun Arya,
  • Srinivasa Narayana,
  • Twinkle Sinha,
  • Samantapudi Venkata Satyanarayana Raju

摘要

Fruit fly infestation poses a major threat to cucurbit production worldwide, with the melon fruit fly (Zeugodacus cucurbitae) being one of the most severe due to its widespread distribution and significant yield losses. Accurate identification of Z. cucurbitae remains challenging due to overlapping adult morphological characters within cryptic species complexes and difficulties in identifying immature stages. To address this, we developed a species-specific primer (SSP) assay targeting the mitochondrial cytochrome c oxidase subunit I (COI) gene for precise identification of Z. cucurbitae. The SSP was subjected to various tests, including cross-specificity, sensitivity densitometric, and real-time PCR assay. The SSP (ZCRP1F/ZCRP1R) exhibited 100% accuracy across specimens from diverse geographic regions in India and demonstrated high specificity, with no cross-amplification among other economically important fruit fly species. Sensitivity tests confirmed detection limits as low as 10 pg/µl in standard PCR and 1 pg/µl in real-time PCR, yielding a 294 bp amplicon. Furthermore, compatibility with the Real Gene Tissue-Direct PCR kit enabled direct amplification from insect tissues, eliminating the need for DNA extraction. This streamlined approach enhances diagnostic efficiency, reducing processing time. The developed assay has significant potential for applications in quarantine inspections, border security, and for prevention of entry, contributing to improved pest management and biosecurity measures.