Investigation of Preanalytical Variables Impacting Pathogen Cell-Free DNA in Blood and Urine
Autor: | Adithya Cattamanchi, Niaz Banaei, Alfred Andama, Catherine A. Hogan, Jason R. Andrews, Damian Madan, Benjamin A. Pinsky, Grant Theron, Amy Steadman, Akos Somoskovi, Zaida Palmer, Byron W.P. Reeve, Malaya K. Sahoo, Kanagavel Murugesan, Julio Croda |
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Přispěvatelé: | Carroll, Karen C, Graduate School |
Rok vydání: | 2019 |
Předmět: |
Male
0301 basic medicine Preservative Urine Medical and Health Sciences Aspergillus fumigatus 0302 clinical medicine Medicine Viral DNA Fungal Pathogen Blood Specimen Collection biology Bacterial Middle Aged Biological Sciences Healthy Volunteers Body Fluids 3. Good health Fungal PCR Infectious Diseases Cell-free fetal DNA 030220 oncology & carcinogenesis Viruses Female Infection Cell-Free Nucleic Acids DNA Bacterial Adult Microbiology (medical) Microbiology Specimen Handling cell-free DNA Vaccine Related Andrology Mycobacterium tuberculosis Young Adult 03 medical and health sciences Rare Diseases Clinical Research Biodefense preanalytical Humans Tuberculosis Liquid biopsy Bacteria liquid biopsy Agricultural and Veterinary Sciences business.industry Prevention Fungi Bacteriology DNA biology.organism_classification Emerging Infectious Diseases Good Health and Well Being 030104 developmental biology DNA Viral Blood Collection Tube business |
Zdroj: | Journal of clinical microbiology, vol 57, iss 11 Journal of clinical microbiology, 57(11). American Society for Microbiology Journal of Clinical Microbiology |
ISSN: | 1098-660X 0095-1137 |
Popis: | Pathogen cell-free DNA (pcfDNA) in blood and urine is an attractive biomarker; however, the impact of preanalytical factors is not well understood. Blood and urine samples from healthy donors spiked with cfDNA from Mycobacterium tuberculosis, Salmonella enterica, Aspergillus fumigatus, and Epstein-Barr virus (EBV) and samples from tuberculosis patients were used to evaluate the impact of blood collection tube, urine preservative, processing delay, processing method, freezing and thawing, and sample volume on pcfDNA. Pathogen cell-free DNA (pcfDNA) in blood and urine is an attractive biomarker; however, the impact of preanalytical factors is not well understood. Blood and urine samples from healthy donors spiked with cfDNA from Mycobacterium tuberculosis, Salmonella enterica, Aspergillus fumigatus, and Epstein-Barr virus (EBV) and samples from tuberculosis patients were used to evaluate the impact of blood collection tube, urine preservative, processing delay, processing method, freezing and thawing, and sample volume on pcfDNA. The PCR cycle threshold (CT) was used to measure amplifiable cfDNA. In spiked samples, the median CT values for M. tuberculosis, S. enterica, and EBV cfDNA were significantly lower in blood collected in K2EDTA tubes than those in Streck and PAXgene blood collection tubes, and they were was significantly lower in urine preserved with EDTA (EDTA-urine) than in urine preserved with Streck reagent (Streck-urine). Blood and urine samples from TB patients preserved with K2EDTA and Tris-EDTA, respectively, showed significantly lower median M. tuberculosis CT values than with the Streck blood collection tube and Streck urine preservative. Processing delay increased the median pathogen CT values for Streck and PAXgene but not K2EDTA blood samples and for urine preserved with Streck reagent but not EDTA. Double-spin compared with single-spin plasma separation increased the median pathogen CT regardless of blood collection tube. No differences were observed between whole urine and supernatant and between fresh and thawed plasma and urine after 24 weeks at –80°C. Larger plasma and urine volumes in contrived and patient samples showed a significantly lower median M. tuberculosis CT. These findings suggest that large-volume single-spin K2EDTA-plasma and EDTA-whole urine with up to a 24-h processing delay may optimize pcfDNA detection. |
Databáze: | OpenAIRE |
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