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Test Code CHIKS Chikungunya Virus, PCR, Molecular Detection, Serum


Additional Testing Requirements


Due to the short period in which chikungunya RNA may be detected in serum, testing serum for IgM- and IgG-class antibodies to chikungunya virus is also recommended. See CHIKV / Chikungunya IgM and IgG, Antibody, Serum.

 

Testing for RNA or IgM-class antibodies to dengue and Zika viruses, concurrently with chikungunya virus testing should also be considered, given the overlapping clinical syndromes seen with these viruses. See the following:

DENGM / Dengue Virus Antibody, IgG and IgM, Serum

DENVP / Dengue Virus Antibody/Antigen Panel, Serum

DENGS / Dengue Virus, Molecular Detection, PCR, Serum

VZIKM / Zika Virus, IgM Antibody Capture ELISA, Serum

VZIKS / Zika Virus, PCR, Molecular Detection, Serum



Specimen Required


Collection Container/Tube:

Preferred: Serum gel

Acceptable: Red top

Submission Container: Sterile container

Specimen Volume: 0.5 mL Serum

Collection Instructions: Within 2 hours of collection, centrifuge and aliquot the serum into a sterile container.

Additional Information: Serum specimens not aliquoted from the serum gel collection tube into a sterile container will be rejected.


Useful For

Qualitative detection of chikungunya virus in serum after early symptom onset (ideally <7 days)

 

This test is not recommended for screening healthy patients.

Reporting Name

Chikungunya Virus, PCR, Serum

Specimen Type

Serum

Specimen Minimum Volume

Serum: 0.3 mL

Specimen Stability Information

Specimen Type Temperature Time
Serum Refrigerated (preferred) 7 days
  Frozen  7 days

Reject Due To

Gross hemolysis Reject
Heat-inactivated specimen Reject

Reference Values

Negative

Reference values apply to all ages.

Supportive Data

The following validation data supports the use of this assay for clinical testing.

 

Accuracy/Diagnostic Sensitivity and Specificity:

Clinical specimens, commercial samples, and a blinded panel of positive and negative samples provided by the Centers for Disease Control and Prevention (CDC) were used for the accuracy experiments. Testing was completed per the manufacturer's instructions, using the easy Mag (bioMerieux) for RNA extraction.

-Ninety-five clinical serum specimens received from the Mayo Infectious Disease Serology Laboratory (IDS) were tested using the Altona RealStar (ARS) Chikungunya (CHIK) RT-PCR Kit 2.0 assay. These specimens had been submitted through Mayo Clinic Laboratories and sent to an external laboratory for serology (IgM IFA) and reverse transcription polymerase chain reaction (RT-PCR) testing in 2014.

-Sixteen vials of human plasma from donor units collected in Puerto Rico extracted using the QIAGEN QIAamp Viral RNA Mini Kit.

-Spiking studies: To supplement the results, negative cerebrospinal fluid (CSF) specimens were spiked with viral RNA of CHIK and tested in a blinded fashion. The spiking material was heat inactivated (HI) CHIK culture fluid (CF).

 

The ARS CHIK results were compared to the consensus results of the ARS CHIK, a published assay by Lanciotti(1) and a commercial assay, the Bio-Rad ZDC Multiplex RT-PCR Assay (ZDC). The gold standard was considered the consensus between 2 of the 3 PCR assays.

 

Results:

The ARS CHIK RT-PCR Kit detected 9 more chikungunya virus-positive specimens from the patient samples received from IDS than the Lanciotti and ZDC RT-PCR assays.

-Eight of the nine ARS CHIK+/Lanciotti-/ZDC- results were positive by chikungunya IgM enzyme immunoassay and/or chikungunya RT-PCR at another commercial reference lab and, therefore, considered likely true positives. Final consensus sensitivity and specificity were 100% and 98% respectively

-There was 100% agreement with among the ARS CHIK, Lanciotti, and CDC Trioplex RT-PCR assays using a CDC validation panel.

-The CSF specimens spiked with HI chikungunya virus CF at low concentrations gave expected results in 44/44 specimens.

 

Analytical Specificity:

No cross-reactivity was observed with the ARS CHIK RT-PCR Kit when tested against a comprehensive specificity panel, which included 32 bacterial, fungal, and viral organisms from culture collections along with well characterized laboratory strains causing similar disease states, closely related organisms, or from organisms commonly found in the specimens tested. This included West Nile virus (lineage 1 and 2), dengue virus (types 1, 2, 3, and 4), tick-borne encephalitis virus, yellow fever virus, Japanese encephalitis virus, Zika virus, and poliovirus. The manufacturer tested additional organisms as listed in the package insert.

Method Description

The RealStar Chikungunya Virus RT-PCR Kit was developed by Altona Diagnostics to assist in the diagnosis of chikungunya (CHIK) infection by testing serum for CHIK RNA. The Altona RealStar Chikungunya (ARS CHIK) RT-PCR Kit 2.0 is a qualitative reverse transcription-PCR (RT-PCR) assay targeting the nonstructural protein 1 (NS1) gene. The assay includes a heterologous amplification system (internal control: IC) to identify possible RT-PCR inhibition and to confirm the integrity of the reagents of the kit. Testing will be performed per the manufacturer's instructions using the LightCycler 480 following extraction with the NucliSENS easyMag (bioMerieux).

 

Real-time RT-PCR technology utilizes a reverse-transcriptase reaction to convert RNA into complementary DNA, PCR for the amplification of specific target sequences, and target specific probes for the detection of the amplified DNA. The probes are labelled with fluorescent reporter and quencher dyes. Probes specific for CHIK RNA are labelled with the fluorophore FAM. The probe specific for the IC is labeled with the fluorophore JOE. Using probes linked to distinguishable dyes enables the parallel detection of CHIK specific RNA and the IC in corresponding detector channels of the real-time PCR instrument.(Package insert: RealStar Chikungunya RT-PCR Kit 2.0. Altona Diagnostics; 01/2017)

Day(s) Performed

Monday, Wednesday, Friday

Report Available

1 to 5 days

Specimen Retention Time

7 days

Performing Laboratory

Mayo Clinic Laboratories in Rochester

CPT Code Information

87798

LOINC Code Information

Test ID Test Order Name Order LOINC Value
CHIKS Chikungunya Virus, PCR, Serum 81152-1

 

Result ID Test Result Name Result LOINC Value
603833 Chikungunya Virus, PCR, Serum 81152-1

Testing Algorithm

For more information see Mosquito-borne Disease Laboratory Testing.

Forms

If not ordering electronically, complete, print, and send a Microbiology Test Request (T244) with the specimen.