title: Opportunities to improve storage and transportation of blood specimens for CD4 testing in a rural district in Zimbabwe using BD vacutainer CD4 stabilization tubes: a stability and diagnostic accuracy study.
abstract: BACKGROUND
  CD4+ T-cell testing of blood specimens collected in standard EDTA Vacutainer tubes and transported at ambient temperature, must be completed within 48 hours with the BD FACSCount™ flow cytometer, restricting specimen collection in remote clinics with no on-site testing and limited specimen transport services. We conducted a study in Buhera District, Zimbabwe, to assess the stability and accuracy of CD4+ T-cell results of samples collected in Stabilization Tubes (ST) and stored at ambient temperature for varying time periods.
  
  
  METHODS
  Paired EDTA and ST samples were collected from 51 HIV-positive patients aged 18 years and older. CD4+ T-cell testing was done on arrival in the laboratory (Day 0). ST samples were retested on Days 3, 5, and 7. Nineteen ST samples were stored for an additional week and retested on Day 14.
  
  
  RESULTS
  There was a strong correlation between absolute CD4+ T-cell counts measured in the EDTA Day 0 reference sample and Day 7 ST sample (Spearman's rho: 0.9778; mean difference: -4.9 cells/μL and limits of agreement (LOA): 98.5 and 88.7 cells/μL); and the reference sample and Day 14 ST sample (Spearman's rho: 0.9632; mean difference 5.1 cells/μL and LOA: -99.6 and 109.8 cells/μL. Using a 350 cells/μL threshold, the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were all 100% on Day 7, and 83.3%, 100%, 100% and 92.9% on Day 14. Using a 500 cells/μL threshold, the sensitivity, specificity, PPV and NVP were 100%, 88.5%, 88.5% and 100% on Day 7 and 88.9%, 80.0%, 80.0% and 88.9% on Day 14.
  
  
  CONCLUSIONS
  CD4 ST can be used and stored up to 7 days as a reliable alternative to standard EDTA tubes in settings where CD4+ T-cell testing within 48 hours is not feasible. Despite the small sample size, results suggest that ST may be stored up to 14 days at room temperature for CD4 testing, without compromising accuracy. However, further studies with larger sample sizes are needed to confirm this preliminary finding.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Fajardo
authors.forename: Carol
authors.initials: C
authors.lastname: Metcalf
authors.forename: Elton
authors.initials: E
authors.lastname: Mbofana
authors.forename: Charlotte
authors.initials: C
authors.lastname: van Vyve
authors.forename: Dhodho
authors.initials: D
authors.lastname: Munyaradzi
authors.forename: Sandra
authors.initials: S
authors.lastname: Simons
authors.forename: Misheck
authors.initials: M
authors.lastname: Kuhudzayi
authors.forename: Helen
authors.initials: H
authors.lastname: Bygrave
mesh_terms: D000328:Adult; D000925:Anticoagulants / Q000737:chemistry; D001800:Blood Specimen Collection / Q000295:instrumentation* / Q000379:methods / Q000592:standards; D018791:CD4 Lymphocyte Count / Q000295:instrumentation / Q000379:methods / Q000592:standards; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D004492:Edetic Acid / Q000737:chemistry; D005260:Female; D005434:Flow Cytometry; D015658:HIV Infections / Q000097:blood* / Q000175:diagnosis / Q000276:immunology; D006801:Humans; D008297:Male; D008875:Middle Aged; D058996:Quality Improvement; D012424:Rural Population; D012680:Sensitivity and Specificity; D014186:Transportation; D015030:Zimbabwe
chemical_list: D000925:Anticoagulants; D004492:Edetic Acid