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
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
Externí odkaz: