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A high-throughput detection method for invasive fruit fly (Diptera: Tephritidae) species based on microfluidic dynamic array

Jiang, Fan ; Fu, Wei ; Clarke, Anthony R. ; Schutze, Mark Kurt ; Susanto, Agus ; Zhu, Shuifang ; Li, Zhihong

Molecular ecology resources, 2016-11, Vol.16 (6), p.1378-1388 [Periódico revisado por pares]

England: Blackwell Publishing Ltd

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  • Título:
    A high-throughput detection method for invasive fruit fly (Diptera: Tephritidae) species based on microfluidic dynamic array
  • Autor: Jiang, Fan ; Fu, Wei ; Clarke, Anthony R. ; Schutze, Mark Kurt ; Susanto, Agus ; Zhu, Shuifang ; Li, Zhihong
  • Assuntos: Anastrepha ; Animals ; Bactrocera ; biological invasion ; Ceratitis ; Dacus ; Diptera ; DNA barcodes ; DNA Barcoding, Taxonomic - methods ; Electron Transport Complex IV - genetics ; Entomology - methods ; Fluidigm system ; Microfluidics - methods ; molecular identification ; plant quarantine ; Plants - parasitology ; Quarantine ; Real-Time Polymerase Chain Reaction ; Rhagoletis ; species-specific ; Tephritidae ; Tephritidae - genetics ; Tephritidae - growth & development ; Time Factors
  • É parte de: Molecular ecology resources, 2016-11, Vol.16 (6), p.1378-1388
  • Notas: istex:BFA8E81CB5F538FAA162705B2A4DA38AC044F813
    ArticleID:MEN12542
    National Science and Technology Support Program of China - No. 2012BAK11B01
    Fig. S1. Morphological characteristics figures of 35 fruit fly species.Fig. S2. Specificity of each species-specific primer pair based on PCR.Fig. S3. Specificity of each species-specific probe and primer set based on real-time PCR.Fig. S4. The COI barcodes alignment of 27 fruit fly species with the placement of primers and probes.Table S1. Information of fruit fly species and populations used in this study.Table S2. Sequence information used for primer and probe design.Table S3. Fruit fly species in their respective assay and sample inlets on the chip.
    Public Welfare Project for Quality Supervision, Inspection and Quarantine - No. 201310091
    ark:/67375/WNG-59RP6LJF-S
    ObjectType-Article-1
    SourceType-Scholarly Journals-1
    ObjectType-Feature-2
    content type line 23
  • Descrição: Invasive species can be detrimental to a nation's ecology, economy and human health. Rapid and accurate diagnostics are critical to limit the establishment and spread of exotic organisms. The increasing rate of biological invasions relative to the taxonomic expertise available generates a demand for high‐throughput, DNA‐based diagnostics methods for identification. We designed species‐specific qPCR primer and probe combinations for 27 economically important tephritidae species in six genera (Anastrepha, Bactrocera, Carpomya, Ceratitis, Dacus and Rhagoletis) based on 935 COI DNA barcode haplotypes from 181 fruit fly species publically available in BOLD, and then tested the specificity for each primer pair and probe through qPCR of 35 of those species. We then developed a standardization reaction system for detecting the 27 target species based on a microfluidic dynamic array and also applied the method to identify unknown immature samples from port interceptions and field monitoring. This method led to a specific and simultaneous detection for all 27 species in 7.5 h, using only 0.2 μL of reaction system in each reaction chamber. The approach successfully discriminated among species within complexes that had genetic similarities of up to 98.48%, while it also identified all immature samples consistent with the subsequent results of morphological examination of adults which were reared from larvae of cohorts from the same samples. We present an accurate, rapid and high‐throughput innovative approach for detecting fruit flies of quarantine concern. This is a new method which has broad potential to be one of international standards for plant quarantine and invasive species detection.
  • Editor: England: Blackwell Publishing Ltd
  • Idioma: Inglês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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