Zika, Chikungunya, and Dengue Viruses: Detection in Mosquitoes, RT-PCR

Laboratory Fee Schedule
Procedure #: BTA0002
CPT: 87040, 87070, 87077


Synonym(s):	Zika virus, ZIKV, flavivirus, Chikungunya virus, CHIKV, alphavirus, Dengue viruses, DENV, arbovirus, mosquito, reverse transcriptase-polymerase chain reaction, RT-PCR, virus isolation, cell culture, indirect fluorescent antibody test, IFA
Requisition Form	G-14
Test Description	Real-time RT-PCR multiplex assay for detection of Zika virus, Chikungunya virus, and Dengue viruses in mosquito samples.
Pre-Approval Needed	Contact the Arbovirus/Entomology Team at ArbovirusEntomologyTeam@dshs.texas.gov to obtain G- 14 forms and the required shipping materials for submitting mosquito samples to the DSHS Laboratory. Zika, Chikungunya, and Dengue testing for mosquitoes is not provided for routine mosquito testing, but it can be provided for certain situations including case investigations and outbreak situations. Contact the Arbovirus/Entomology Team at ArbovirusEntomologyTeam@dshs.texas.gov for more information.
Supplemental Information Required	N/A
Supplemental Form(s)	N/A
Performed on Specimens from (sources)	Non-human Mosquitoes
Sample/Specimen Type for Testing	Live, adult mosquitoes
Minimum Volume/Size Required	1 (one)mosquito
Storage/Preservation Prior to Shipping	Use a handheld aspirator to transfer up to 100 live mosquitoes from each trap collection to a DSHS Lab- provided plastic shipping carton. Submit only one carton per trap. Store at room temperature until shipment. Do not freeze mosquitoes. Ship mosquitoes as soon as possible after collection.
Transport Medium	N/A
Specimen Labeling	Each shipping carton must be submitted with a completed G-14 submission form. Attach submission form to carton using a rubber band. Ensure collection data is provided in submission form.
Shipping and Specimen Handling Requirements	Ship mosquitoes cool to the Laboratory using the insulated boxes provided by the Arbovirus/Entomology Laboratory. Include provided coolant bottle in the shipment. Fill bottle with water to the line, tighten cap, and freeze. Place frozen bottle in plastic bag (provided) and wrap with damp newspaper or paper towels. Add several sheets of moistened paper towels or newspaper to the inner box to keep humidity at a high level. Avoid saturating the paper; excess water in cartons will damage mosquitoes. Tape the shipping box closed and affix required labels to exterior of the box (boxes are reused). If replacement or additional shipping supplies are needed, contact the Arbovirus/Entomology Team by emailing ArbovirusEntomologyTeam@dshs.texas.gov. Ship for delivery to Lab during business hours only (Monday to Friday 8 a.m. – 5 p.m.) . Do not ship on Fridays. Shipping overnight or next day is preferred. Additional arbovirus field surveillance techniques are available online here.
Method	Real-time RT-PCR multiplex assay.
Turn-around Time	5 days
Interferences/Limitations	N/A
Common Causes for Rejection	Incorrect storage conditions of mosquitoes. Species identification of mosquitoes received in poor condition may not be possible. Mosquitoes that cannot be identified to the species level will not be tested. Non-vector species submitted. Only vector species are tested.
Additional Information	A Dengue virus multiplex RT-PCR assay is conducted as a reflex test to confirm Dengue detection and to determine the serotype (DENV 1 –4). This dengue virus multiplex testing cannot be ordered by submitters. Additional arbovirus testing using cell culture screening methods is conducted on 20 percent of submitted mosquito trap collections. This testing cannot be ordered by submitters. Arbovirus identification using an indirect fluorescent antibody test is conducted as a reflex test when cytopathic effects are detected during cell culture screening. This testing cannot be ordered by submitters.