AIDS Rev. 2014;16:160-71 (Supplementary Data)
Measurement of HIV-1 Viral Load for Drug Resistance Surveillance using Dried Blood Spots: Literature Review and Modeling of Contribution of DNA and RNA
Neil T. Parkin
Table 1. Comparison table
First author
Year
published
Journal or Conference
Title
PubMed ID
Method
Patients on ART
VL range
(log c/ml
in plasma)
N paired
specimens
N < 1,000
in plasma
Fiscus
1998
J Clin Microbiol
Quantitation of human immunodeficiency virus type 1 RNA in plasma by using
blood dried on filter paper
9431960
NASBA
Not reported
Uttayamakul
2005
J Virol Methods
Usage of dried blood spots for molecular diagnosis and monitoring HIV-1 infection
15913797
Boom extraction + NASBA
AlvarezMunoz
2005
Arch Med Res
High correlation of human immunodeficiency virus type-1 viral load measured in
dried-blood spot samples and in plasma under different storage conditions
15950079
Ayele
2007
J Clin Microbiol
Use of dried spots of whole blood, plasma, and mother’s milk collected on filter
paper for measurement of human immunodeficiency virus type 1 burden
Waters
2007
JAIDS
Kane
2008
Leelawiwat
N < LOD
in plasma
<3–>6
76
17
0
No
<2–>5
30
6
0?
Nuclisens extraction, quantitation by EasyQ (DBS) and
Roche Amplicor (plasma)
Not reported
<2–>5
108
17251400
Primagen Retina Rainbow
Not reported
2.5 – 6
Evaluation of Filter Paper Transfer of Whole-Blood and Plasma Samples for
Quantifying HIV RNA in Subjects on Antiretroviral Therapy in Uganda
18193501
Primagen extraction/Roche CTM (DBS) vs. Roche
Amplicor (plasma)
Yes
J Virol Methods
Quantitation of HIV-1 RNA in dried blood spots by the real-time NucliSENS EasyQ
HIV-1 assay in Senegal
18242718
Nuclisens extraction, quantitation by EasyQ
Not reported (assumed to be a mixture)
2009
J Virol Methods
Dried blood spots for the diagnosis and quantitation of HIV-1: stability studies and
evaluation of sensitivity and specificity for the diagnosis of infant HIV-1 infection in
Thailand
18952125
Nuclisens extraction, quantitation by EasyQ
Monleau
2009
J Clin Microbiol
Evaluation of Different RNA Extraction Methods and Storage Conditions of Dried
Plasma or Blood Spots for Human Immunodeficiency Virus Type 1 RNA
Quantification and PCR Amplification for Drug Resistance Testing
19193835
Monleau
2009
J Clin Microbiol
Evaluation of Different RNA Extraction Methods and Storage Conditions of Dried
Plasma or Blood Spots for Human Immunodeficiency Virus Type 1 RNA
Quantification and PCR Amplification for Drug Resistance Testing
Garrido
2009
J Clin Microbiol
Garrido
2009
Marconi
Specificity
(LOD)
Sensitivity
(LOD)
Specificity
(1,000)
Sensitivity
(1,000)
Specificity
(1,000)
< 90%
Sensitivity
(1,000)
< 90%
DBS false
positive
DBS false
positive
Comment
4
Nr
90%
Few (4?)
Few (4?)
5
6
0
0
65
2
© Permanyer Publications 2014
Data First Consulting, Belmont, California, USA
1
0
306
281
278
2.9 – 5.4
41
9
6
0
2
No
3 – >5
56
0
0
0
0
Abbott extraction; quantitation by Biocentric Generic
assay
Not reported (assumed no)
<2 – >5
39
2
0
High
High
1
1
DBS LOQ ~3.7 log
19193835
Nuclisens extraction; quantitation by Biocentric Generic
assay
Not reported (assumed no)
<2 – >5
39
2
0
High
High
1
0
DBS LOQ ~3.7 log
Correlation between Human Immunodeficiency Virus Type 1 (HIV-1) RNA
Measurements Obtained with Dried Blood Spots and Those Obtained with Plasma
by Use of Nuclisens EasyQ HIV-1 and Abbott RealTime HIV Load Tests
19193847
Abbott RealTime HIV
Not reported (assumed to be a
mixture)
<2–>5
81
11
0?
High
76%
0
19
DBS LOD ~3.72 log
J Clin Microbiol
Correlation between Human Immunodeficiency Virus Type 1 (HIV-1) RNA
Measurements Obtained with Dried Blood Spots and Those Obtained with Plasma
by Use of Nuclisens EasyQ HIV-1 and Abbott RealTime HIV Load Tests
19193847
Nuclisens extraction, quantitation by EasyQ
Not reported (assumed to be a
mixture)
<2–>5
97
12
0
High
84%
0
13
DBS LOD ~3.48 log
2009
Clin Micro Inf
Evaluation of the Abbott Real-Time HIV-1 quantitative assay with dried blood spot
specimens
19220340
Abbott RealTime HIV
Not reported (assumed no)
2.3 – > 5
168
20
0
Na
97%
0
5
Ikomey
2009
J Int Assoc Physicians AIDS
Care (Chic)
Dried blood spots versus plasma for the quantification of HIV-1 RNA using the
manual (PCR-ELISA) amplicor monitor HIV-1 version 1.5 assay in Yaounde,
Cameroon
19357423
Roche Amplicor
No
2.5 – 6
60
7
3
3
2
Mehta
2009
PLoS One
Low-cost HIV-1 diagnosis and quantification in dried blood spots by real time PCR
19503790
Real-Time LightCycler (rtLC) PCR assay (DBS) vs. bDNA
or Amplicor (plasma)
51%
<2–>5
37
9
5
100%
92%
89%
100%
1
0
0
3
Reigadas
2009
J Virol Methods
Quantitation of HIV-1 RNA in dried blood and plasma spots
19523984
Qiagen extraction; quantitation by CTM for plasma,
Biocentric for DBS
20%
<2–7
68
15
7
57%
85%
67%
92%
1
0
3
9
DBS LOD 2.87 log
Mbida
2009
JAIDS
Measure of viral load by using the Abbott Real-Time HIV-1 assay on dried blood
and plasma spot specimens collected in 2 rural dispensaries in Cameroon.
19620878
Abbott RealTime HIV
42% of series 1 (all <40 in plasma)
<2–>5
45
17
9
100%
83%
100%
100%
0
0
0
0
DBS LOD 3 log; FP/FN using 1,000 threshold, specificity at 1,000 inferred
Johannessen
2009
Clin Infect Dis
Dried blood spots perform well in viral load monitoring of patients who receive
antiretroviral treatment in rural Tanzania.
19663598
Nuclisens extraction, quantitation by EasyQ (DBS); Roche
CTM or Amplicor (plasma)
yes
<2–>5
98
61
33
94%
54%
97%
76%
0
1
2
9
DBS LOD ~3.48 log
Lofgren
2009
AIDS
Evaluation of a dried blood spot HIV-1 RNA program for early infant diagnosis and
viral load monitoring at rural and remote healthcare facilities
19741481
Abbott RealTime HIV (all) and Roche CAP/CTM (subset)
53%
<2–>5
137
55
87%
99%
7
1
DBS LOD ~2.6 log; spec/sens threshold 400; numbers uncertain, cannot
calculatel sens/spec at 1,000
Andreotti
2010
J Clin Virol
Correlation between HIV-1 viral load quantification in plasma, dried blood spots,
and dried plasma spots using the Roche COBAS Taqman assay
19962936
Nuclisens extraction, quantitation by Roche CTM
17% (15% of specimens)
<2–>5
129
34
19
95%
88%
97%
96%
0
0
1
13
DBS LOQ 2.5 log
van Deursen
2010
J Clin Virol
Measuring human immunodeficiency virus type 1 RNA loads in dried blood spot
specimens using NucliSENS EasyQ HIV-1 v2.0
20018560
Nuclisens extraction, quantitation by EasyQ
Yes
<2–>5
224
153
94
77%
81%
100%
80%
0
1
0
14
DBS LOD ~2.9 log; FP/FN using 1,000 threshold
Monleau
2010
J Antimicrob Chemother
Effect of storage conditions of dried plasma and blood spots on HIV-1 RNA
quantification and PCR amplification for drug resistance genotyping
20542904
Nuclisens extraction; quantitation by Biocentric Generic
assay
Not reported (same specimens as
2009 jcm paper)
< 2 – >5
34
Viljoen
2010
J Acquir Immune Defic
Syndr
Dried blood spot HIV-1 RNA quantification using open real-time systems in South
Africa and Burkina Faso
20700058
Nuclisens extraction; quantitation by Biocentric Generic
assay
Mixed (mostly not)
2.5 – > 6
327
15
0
Viljoen
2010
J Acquir Immune Defic
Syndr
Dried blood spot HIV-1 RNA quantification using open real-time systems in South
Africa and Burkina Faso
20700058
Nuclisens extraction; quantitation by Biocentric Generic
assay
Yes
<2–>5
28
21
18
Balamane
2010
Open Virol J
Detection of HIV-1 in Saliva: Implications for Case-Identification, Clinical Monitoring
and Surveillance for Drug Resistance
21673840
Nuclisens extraction, quantitation by Roche Amplicor 1.5
Not reported
<2–>5
32
6
5
Pirillo
2011
J Antimicrob Chemother
Quantification of HIV-RNA from dried blood spots using the Siemens VERSANT®
HIV-1 RNA (kPCR) assay
21930572
Siemens kPCR
Not reported
2–7
98
26
13
69%
88%
Arredondo
2012
J Clin Microbiol
Comparison of HIV-1 RNA Measurements Obtained by Using Plasma and Dried
Blood Spots in the Automated Abbott Real-Time Viral Load Assay
22170904
Abbott RealTime HIV
30%
<2–>5
154
26
57
91%
77%
Vidya
2012
J Virol Methods
Dried blood spots versus plasma for the quantitation of HIV-1 RNA using a
real-Time PCR, m2000rt assay
22401801
Abbott RealTime HIV
Not reported (assumed no)
<2–>5
100
13
High
87%
Rottinghaus
2012
Clin Infect Dis
Dried blood spot specimens are a suitable alternative sample type for HIV-1 viral
load measurement and drug resistance genotyping in patients receiving first-line
antiretroviral therapy
22412066
Nuclisens extraction, quantitation by EasyQ
Yes
<2–>5
173
155
147
David
2012
Indian J Med Microbiol
Comparison of HIV-1 RNA level estimated with plasma and DBS samples: a pilot
study from India (South)
23183463
Abbott RealTime HIV
Mixed
3.5 – >6
52
10
10
Ouma
2013
J Clin Microbiol
Evaluation of Quantification of HIV-1 RNA Viral Load in Plasma and Dried Blood
Spots by Use of the Semiautomated Cobas Amplicor Assay and the Fully
Automated Cobas Ampliprep/TaqMan Assay, Version 2.0, in Kisumu, Kenya
23390278
Roche CAP/CTM
No (baseline before PMTCT)
< 2 – >5
421
28
Neogi
2012
Indian J Med Res
Dried blood spot HIV-1 RNA quantification: a useful tool for viral load monitoring
among HIV-infected individuals in India
23391790
Abbott RealTime HIV
Mixed
<2–>6
120
49
Aitken
2013
J Clin Microbiol
Development and evaluation of an affordable real-time qualitative assay for
determining HIV-1 virological failure in plasma and dried blood spots
23596235
ARTA-VFA
Mixed
<2–>6
82
Yapo
2013
J Virol Methods
Evaluation of dried blood spot diagnosis using HIV1-DNA and HIV1-RNA Biocentric
assays in infants in Abidjan, Cote d’Ivoire. The Pedi-Test DBS ANRS 12183 Study
23872283
Qiagen extraction; quantitation by Biocentric
No
3–>5
46
1
MercierDelarue
2014
J Clin Microbiol
Higher Specificity of NASBA Isothermal Technology Versus Real-Time PCR for
HIV-1 RNA Quantification on Dried Blood Spots
24131691
Nuclisens extraction, quantitation by EasyQ
Mixed
<2–>5
193
97
98%
89%
0
1
2
11
Threshold used 800 copies/ml
MercierDelarue
2014
J Clin Microbiol
Higher Specificity of NASBA Isothermal Technology Versus Real-Time PCR for
HIV-1 RNA Quantification on Dried Blood Spots
24131691
Roche CAP/CTM
Mixed
<2–>5
196
70
83%
97%
1
0
12
4
Threshold used 800 copies/ml
Monleau
2014
J Clin Microbiol
Dried Blood Spots (DBS) for HIV-1 viral load and Drug Resistance monitoring in pa)
ents receiving HAART in Africa and Asia: The ANRS 12235 study
24478491
Nuclisens extraction, quantitation by EasyQ
Yes
<2–>5
91
24
96%
85%
0
1
1
10
Also presented at croi 2013 (ayouba et al)
Monleau
2014
J Clin Microbiol
Dried Blood Spots (DBS) for HIV-1 viral load and Drug Resistance monitoring in pa)
ents receiving HAART in Africa and Asia: The ANRS 12235 study
24478491
Abbott RealTime HIV
Yes
<2–>5
173
124
93%
94%
0
0
9
3
Also presented at croi 2013 (ayouba et al)
Monleau
2014
J Clin Microbiol
Dried Blood Spots (DBS) for HIV-1 viral load and Drug Resistance monitoring in pa)
ents receiving HAART in Africa and Asia: The ANRS 12235 study
24478491
Nuclisens extraction; quantitation by Biocentric Generic
assay
Yes
<2–>5
118
79
61%
92%
1
0
31
3
Also presented at croi 2013 (ayouba et al)
Fajardo
2014
J Clin Microbiol
Prospective Evaluation of the Diagnostic Accuracy of Dried Blood Spots from
Finger-Prick for the Determination of HIV-1 Viral Load with the NucliSENS Easy-Q
HIV-1 v2.0 in Malawi
24501032
Nuclisens extraction, quantitation by EasyQ
Yes
<2–>5
610
508
97%
91%
0
0
14
MtapuriZinyowera
2013
IAS
Field evaluation of performance of dried blood spots (DBS) from finger-prick for the
determination of viral load in a resource-constrained setting in urban and rural
Zimbabwe
na
Roche CAP/CTM
Mixed
<2–>5
118
76
25%
97%
1
0
57
Rutstein SE
2014
J Clin Virol
Measures of viral load using Abbott RealTime HIV-1 Assay on venous and
fingerstick dried blood spots from provider-collected specimens in Malawian
District Hospitals
24906641
Abbott RealTime HIV
Yes
<2–>5
149
137
97%
100%
0
0
4
77%
90%
91%
98%
1
0
0
0
FP/FN using 1,000 threshold, uncertain of numbers
DBS LOD 2.3 log?
dNAse treatment reduced DBS VL (closer to DPS)
19
93%
17%
100%
26
19%
83%
100%
97%
1
1
0
0
17
0
4
0
4
10
15
2
DBS LOD ~2.74 log; FP/FN using 1,000 threshold, specificity at 1,000
inferred
0
6
DBS LOQ ~3.3 log
128
DNAse treatment reduced DBS false pos (1,000) to 2
DBS LOQ ~3 log
98%
78%
0
1
3
4
FP/FN using 1,000 threshold
VL all >1,000 if detectable; 10 undetectable in plasma and DBS
100%
100%
100%
100%
0
0
0
0
78%
100%
93%
100%
0
0
2
0
84%
100%
97%
0
0
0
17
0
7
0
425
DBS LOQ ~3.2 log; DBS only tested if plasma VL > 300
Used 5,000 threshold; cannot calculate sens/spec @1,000
vL high
Good sensitivity of DBS compared to HIV-1 RNA plasma but very low
specificity, which translated in a higher rate of false positive results with
DBS
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17
0
ART: antiretroviral therapy; VL: viral load; LOD: level of detection; FP/FN: false positive/false negative; DBS: dried blood spot; LOQ: limit of quantitation
1