title: Detecting DNA viruses in oral fluids: evaluation of collection and storage methods.
abstract: Storing saliva for nucleic acid diagnostics is problematic in resource-constrained settings. DNA Genotek's OMNIgene™·DISCOVER kit aims to stabilise microbial DNA at room temperature. We evaluate this for long-term storage, determining DNA quantity/purity and human herpesvirus 8 (HHV-8) load as indicator. Viral loads and DNA degradation were assayed over 14months in HHV-8-negative saliva spiked with cell-associated and cell-free virus and saliva collected fresh frozen and into kits from 10 HIV-positive patients. Viral loads remained constant for 6-9months, yielding high quantities of DNA: subsequent losses were ≤48%. Patient samples, frozen or kit stored, produced pure DNA of comparable concentration. Higher HHV-8 detection in frozen saliva resulted from losses during ethanol precipitation using kits. After 14months, DNA degradation was significant in frozen saliva, but that in kits had integrity similar to fresh samples. Storing frozen saliva is detrimental. This kit is well suited for collection, long-term storage, and assay of viral DNA in resource-constrained settings.
authors: School of Dentistry and Oral Health, Griffith University, Queensland, Australia; Molecular Basis of Disease Research Program, Menzies Health Institute Queensland, Griffith University, Queensland, Australia; Population & Social Health Research Program (Population Oral Health), Menzies Health Institute Queensland, Griffith University, Queensland, Australia. Electronic address: d.speicher@griffith.edu.au.
authors: David J
authors: DJ
authors: Speicher
authors: Centre for Public Health Research, Kenya Medical Research Institute, Nairobi, Kenya.
authors: Peter
authors: P
authors: Wanzala
authors: Centre for Public Health Research, Kenya Medical Research Institute, Nairobi, Kenya.
authors: Melvin
authors: M
authors: D'Lima
authors: Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA.
authors: Karen E
authors: KE
authors: Johnson
authors: Molecular Basis of Disease Research Program, Menzies Health Institute Queensland, Griffith University, Queensland, Australia; Population & Social Health Research Program (Population Oral Health), Menzies Health Institute Queensland, Griffith University, Queensland, Australia.
authors: Newell W
authors: NW
authors: Johnson
mesh_terms: D000293:Adolescent; D000328:Adult; D005260:Female; D006566:Herpesviridae Infections / Q000821:virology*; D019288:Herpesvirus 8, Human / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D011309:Preservation, Biological / Q000379:methods*; D012463:Saliva / Q000821:virology*; D013048:Specimen Handling / Q000379:methods*; D013997:Time Factors; D019562:Viral Load; D055815:Young Adult
keywords: DNA Genotek; DNA integrity; HHV-8; Herpesvirus; Long-term storage; Salivary diagnostics