HHV6 PCR Quant w/reflex to HHV6 Chromosome Integration

The detection and quantitation of Human Herpesvirus 6 (HHV-6) by real-time PCR amplification. A positive HHV-6 quantitation (HH6QN) result will reflex to order HHV-6 chromosomal integration and typing (HH6ABC). Droplet Digital PCR partitions a single bulk reaction into ~20,000 individual reactions that are each read as positive or negative for DNA template to provide an absolute quantitation of DNA copies without the use of a standard curve. The result of HHV-6 copies/cell is resulted as a ratio obtained by (HHV-6 copies)/(RPP30 copies/2). If the HHV-6 virus to cell ratio is approximately “one”, results are consistent with HHV-6 chromosomal integration of the virus into the patient cell genome.

This test was developed and its performance characteristics determined by the University of Washington Department of Laboratory Medicine and Pathology. It has not been cleared or approved by the US Food and Drug Administration.

For HHV-6 Quantitation Testing, see HHV6 Quantitative by PCR [HH6QN].

For HHV-6 Chromosome Integration and Typing, see Human Herpes 6 Virus Chromosome Integration and Typing [HH6ABC].

Whole Blood

Collect 5 mL of whole blood in an EDTA (lavender-top) tube. Whole blood is required for HH6QN and HH6ABC testing.

Send whole blood to the laboratory at room temperature within 24 hours of collection. Do not centrifuge or freeze the specimen.

Note: If no additional plasma or serum was collected, this specimen may also be used for HH6QN testing. Processing will be performed by UW Molecular Virology.

Unacceptable specimens: Green-top, red-top, or gold SST tubes. Specimens that have been centrifuged or frozen will not be accepted.

DNA Specimens (HH6ABC only)

Extracted DNA specimens are acceptable provided the DNA concentration is 50–100 ng/μL and the volume is at least 100 μL. Extracted DNA must be stored at -20°C or colder and transported frozen.

The laboratory accepts isolated or extracted nucleic acid only from CLIA-certified laboratories or laboratories meeting equivalent requirements as determined by CAP.

Outside Laboratories

Send whole blood to the testing laboratory at room temperature whenever possible.

If the specimen cannot be delivered to the testing laboratory at room temperature within 48 hours of collection, it should be shipped refrigerated at 2–8°C using an ice pack. Specimens received more than 48 hours after collection may be rejected if an adequate buffy coat (cellular fraction) cannot be obtained during processing by the testing laboratory.

Send whole blood specimens to the Lab immediately at room temperature.

The Laboratory MUST receive the sample within 48 hours of collection. If sample cannot get to the testing laboratory at room temperature within 48 hours, send whole blood at 2-8°C (ice pack).

Extracted DNA must be stored at -20°C to -80°C and transported to the lab frozen.

Whole Blood: Send whole blood to the Molecular Virology Laboratory immediately at room temperature. The laboratory must receive the specimen within 48 hours of collection. Upon receipt, the Molecular Virology Laboratory will separate the whole blood into plasma and cellular fractions.

DNA Specimens: Extracted DNA specimens are acceptable provided the DNA concentration is 50–100 ng/μL and the volume is at least 100 μL. Extracted DNA must be stored at -20°C to -80°C and transported to the laboratory frozen.

Outside Laboratories: Send whole blood to the testing laboratory at room temperature whenever possible. If the specimen cannot reach the testing laboratory within 48 hours of collection, ship the whole blood refrigerated at 2–8°C with an ice pack.