Superior samples • Proven performance

Case study
High quality microbial DNA collected withOMNIgene®•ORAL enables detection of HHV-8 in saliva
David J. Speicher and Newell W. Johnson

Griffith Health Institute, Griffith University, Queensland, Australia

Study overview“The DNA Genotek system has
been a great asset in the detection
and genotyping of human herpes
viruses in our field studies on DNA
viruses in India and Kenya. The
DNA Genotek collection kits haveallowed us to detect and genotype
HHV-8 from Kenyan HIV-positive
patients 14 months post collection
with the samples stored at room
temperature, which in Kenya canexceed 30°C.”
David J. Speicher and Newell W. Johnson,
Griffith Health Institute, Griffith University,Queensland, Australia

Saliva is a diagnostic specimen for Human Herpes Virus 8 (HHV-8), however
traditional methods involve collection of saliva as a whole mouth fluid (WMF),
freezing immediately after collection and extraction of DNA via spin columns.
These methods can be useful in developed countries but difficult to use in developing
nations where cold storage and lab infrastructure is scarce. Researchers at Griffith
University in Queensland, Australia first studied the detection of cultured HHV-8
in spiked saliva samples under laboratory conditions. The study then expanded
to include an assessment of DNA Genotek’s OMNIgene®•ORAL† product line
as an alternative to the previous standard for use in field collections
for quantification of HHV-8 oral shedding in HIV-positive patients and for
determining HHV-8 subtypes.

Main challenges
Primary challenges for this study included collecting DNA samples in Kenyawith resource poor infrastructures and ambient temperatures that can be as high as
30°C or more. The researchers were also challenged with collecting samples ata great distance from the processing laboratory which was in Australia. A productthat offered easy collection, reliable stabilization and transport without causing any
degradation of samples would be beneficial in environments such as this.

Collection methods considered
Other collection methods included in this study were frozen saliva, collectedby dribbling WMF into a chilled collection cup or via mouthwash, both ofwhich require immediate chilling of the sample to 4°C and then at -80°C for
long-term storage.

† Samples for the study were collected with DNA Genotek prototype kit P-021 as part of the development of the OMNIgene•ORAL
product line for the collection and stabilization of microbial nucleic acids. The first commercially available kit for this productline is the OMNIgene•ORAL OM-501 format.

For research use only,
not for use in diagnostic procedures.
Not available for clinical diagnostic
use in the United States.

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Why OMNIgene•ORALThe Griffith University researchers included OMNIgene•ORAL in the study
as it is designed to collect and stabilize microbial DNA from saliva at ambient
temperature; making it a good choice for field research, especially where cold
storage of samples is difficult.
David J. Speicher

Newell Johnson

Results‡
Samples were collected from 10 HIV-positive patients in Kenya at ambient
temperature using the OMNIgene•ORAL kit. The specimens were processed
14 months later in Australia. The researchers were able to determine thatOMNIgene•ORAL retained a high quality DNA that is ideal for viral sub typing,even after storage for 14 months at room temperature. The high quality microbial
DNA collected using the OMNIgene•ORAL kit enabled the HHV-8 subtypes to be
determined to full length ORFK1 (1.2 Kb). The researchers were also able to detect
the HHV-8 virus using the OMNIgene•ORAL kit from samples stored at roomtemperature and this produced consistent results. The kits produced high quality
DNA with yields as high or higher than frozen WMF. Overall, OMNIgene•ORAL
is a great asset for doing field studies on DNA viruses in developing countries.

‡ The results were presented for the first time at the Australian Society for Microbiology meeting, July 2011.

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All DNA Genotek protocols, white papers and application notes, are available in the support section of our website at www.dnagenotek.com.

Superior samples • Proven performance
MK-00018 Issue 2/2015-04© 2015 DNA Genotek Inc., a subsidiary of OraSure Technologies, Inc., all rights reserved.

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