Description
Material provided: Summary report on Anopheles/ Aedes mosquito genotyping and pathogen detection
Unit definition: Genotyping of one mosquito sample (20-50 female mosquitoes) × 20-30 markers
Monitoring mosquito vector populations is an integral component of control programs and a prerequisite for effective interventions.
By using RNAlater preserved samples (4×10 female mosquitoes per field population/strain), and either multiplex Taqman assays OR our novel Labdisk platform (www.dmc-malvec.eu), we will provide to the user genotyping for approximately 20-30 most relevant to vector control markers.
The use of pools of mosquitoes, in line with bioassays and microarray based assays has been introduced and recommended, although individual mosquitoes can also be genotyped.
Use of the Unit will provide sufficient information for several research questions, related with the presence and frequency of insecticide resistance alleles, mosquito species and presence of pathogens, in certain ecosystems.
Description of the experiment and the product
TaqMan assays are specific and sensitive reliable assays, available for both individual and pools of mosquitoes. We offer here a multiplexed panel of TaqMan assays, performed either in a classical way (real time PCR) or in LabDisk platform.
The user should provide mosquito samples (lab strains or field-caught) appropriately stored in RNA later (attached SOP for sample preservation in Appendix 1). Consultation for the sampling procedures (technical and biological tips, depending on research questions) could also be provided by the Provider.
We will handle and analyze the samples provided by the user with the following workflow:
Step 1: Mechanic homogenization and chemical lysis of mosquito tissue samples
Step 2: DNA and RNA extraction and cDNA synthesis
Step 3: Multiplex TaqMan assays (in tube and/or LabDisk)
Step 4: Analysis of data and preparation of reports to be returned to the user.
In each step, relevant assurance quality procedures and appropriate control will be used.
Data layout
The data that will be returned to the user will consist of both raw data, as well as summary tables and figures, to presence and percentage of, different species/molecular forms, different insecticide resistance alleles, the levels of major detoxification genes (compared to reference samples) and the presence of pathogens. Additionally, raw qPCR data and a quality report of the procedure will be also made available upon request.
Publications
- Vontas J et al, Stud Health Technol Inform. 2016; 224:54-60.
- Czilwik et al., Lab Chip. 2015;15(18):3749-59. doi: 10.1039/c5lc00591d.
- Vontas et al, Trends Parasitol. 2014 Apr;30(4):191-204
- Bass C et al, Malar Res Treat. 2010; 2010:190434
For more information, please contact us.