Effect of storage conditions of dried plasma and blood spots on HIV-1 RNA quantification and PCR amplification for drug resistance genotyping.

Author(s): Monleau M, Butel C, Delaporte E, Boillot F, Peeters M

Publication: J Antimicrob Chemother, 2010, Vol. 65, Page 1562-6

PubMed ID: 20542904 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of storage duration, storage temperature, and DNAse treatment on HIV VL and amplification of RT using DBS and DPS rather than plasma. DPS and DBS were initially stored at -20 degrees C with a silica desiccant for 5 months and then stored for up to 2 (DPS) or 3 (DBS) months at 20 degrees C in standard laboratory air or at 37 degrees C in an incubator with water trays. The authors did not specify if liquid plasma was fresh or frozen.

   Summary of Findings:

   In DBS stored for 5 months at -20 degrees C, the VL were 0.19 log copies/mL higher than those measured in the corresponding liquid plasma specimens. When the same DBS were then stored for an additional 3 months at 20 degrees C, VL were comparable to those found in liquid plasma, but storage of DBS for 2 months or more at 37 degrees C, after the 5 months at -20 degrees C, resulted in slightly lower VL than those found in liquid plasma. After 5 months at -20 degrees C, the VL in DPS were 0.30 log copies/mL higher than in liquid plasma, but this difference was not significant. When DPS were subsequently stored at 20 degrees C, a slight but insignificant decrease in VL was noted after 2 months (longest period examined in DPS); however when DPS were stored at 37 degrees C, after the 5 months at -20 degrees C, a significant decline in VL was observed after 1 or 2 months. DBS had higher VL than DPS, and treatment of DBS with DNAse significantly decreased the VL in DBS. Amplification of the RT gene was possible in all 12 DBS and 10/12 DPS after storage for 5 months at -20 degrees C. After subsequent storage at 20 degrees C, RT was amplified in 11/12 DBS after 1 or 2 months, 10/11 DBS after 3 months, 4/12 DPS after 1 month and 5/12 DPS after 2 months. After storage at 37 degrees C, after the 5 months at -20 degrees C, RT was successfully amplified in 10/12 DBS after 1 month, 7/12 DBS after 2 months, 8/10 DBS after 3 months, and in 0/11 DPS after 1 month and 0/7 DPS after 2 months. Treatment of DBS with DNAse greatly reduced the number of DBS from which RT could be amplified after storage at 20 or 37 degrees C.

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Plasma

   Preservative Types

   • Other Preservative

   Diagnoses:

   • AIDS/HIV-related

   Platform:

   Analyte	Technology Platform
   RNA	RT-PCR
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage temperature	-20 degrees C 20 degrees C 37 degrees C
   Storage	Storage duration	0 months 1 month 2 months 3 months 5 months
   Storage	Storage conditions	With regularly changed silica desiccant In laboratory air (dry) In incubator with water
   Biospecimen Aliquots and Components	Blood and blood products	Plasma Whole blood
   RT-PCR Specific	Targeted nucleic acid	Reverse transcriptase region of pol
   Analyte Extraction and Purification	Nucleic acid digestion	DNAse treated No DNAse treatment

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