Evaluation of the use of dried spots and of different storage conditions of plasma for HIV‐1 RNA quantification

Amellal, Bahia; Katlama, C.; Cálvez, Vincent

doi:10.1111/j.1468-1293.2007.00484.x

Cited by 35 publications

(43 citation statements)

References 12 publications

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“…However, it has been reported that samples can maintain their HIV-1 RNA concentrations for at least 14 days at 5°C and around 3 years at Ϫ20°C (14,16). Amellal and colleagues also demonstrated that 1 week of storage of plasma specimens at 4, 22, and 30°C did not significantly affect HIV-1 RNA viral load levels (3,4). In contrast, they obtained reduced median HIV RNA concentrations for samples stored at 37°C.…”

Section: Discussionmentioning

confidence: 94%

Low Prevalence Rate of Indeterminate Serological Human Immunodeficiency Virus Results among Pregnant Women from Burkina Faso, West Africa

et al. 2010

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In sub-Saharan Africa, rapid testing for human immunodeficiency virus (HIV) is the most efficient and sometimes the only feasible way to quickly provide information about HIV status among adults and children Ն18 months of age (6,29). In contrast to enzyme immunoassays (EIAs) and Western blot assays (WBs), HIV rapid tests are relatively cheap, easy to use, and fast to perform. Most of them do not require refrigeration, sophisticated laboratory equipment, skilled technicians, and an electricity supply. Results from serum, plasma, whole-blood, urine, or saliva samples are obtained by visual reading after a few minutes. Some of the rapid tests can distinguish HIV type 1 (HIV-1) from HIV type 2 (HIV-2). They are also accurate and reliable as a result of applying a quality system approach recommended by the World Health Organization (WHO) (36). Due to their low cost and technical advantages, they have been adopted into national HIV voluntary counseling and testing (VCT) guidelines in many African countries. Their sensitivity and specificity have been studied in Kenya (12), Tanzania (22), Uganda (15), Zambia (30), South Africa (25), Cameroon (1), Central African Republic (24), Democratic Republic of Congo (19), Ghana (2), Ivory Coast (32), and Burkina Faso (23, 28).One objective for rapid HIV testing is to minimize the occurrence of indeterminate results (i.e., discordant results when using at least two different rapid tests). It is often quite difficult for HIV counselors and health care providers to disclose such indeterminate results. In the context of interventions for prevention of mother-to-child transmission (PMTCT) of HIV-1 (which can require immediate decisions, notably during labor), medical staff need to know rapidly the HIV status of the woman from the laboratory in order to provide her, or not, antiretroviral (ARV) prophylaxis (5,26,27). The prolonged delay in the decision (by performing additional tests or by retesting women 14 days later) can be inappropriate in the clinical routine since women may deliver before obtaining definitive results.The aims of this study conducted among pregnant women from Burkina Faso (West Africa) with a low risk of early HIV-1 seroconversion were (i) to determine the prevalence rates of indeterminate results by using two rapid tests in a sequential algorithm, as recommended in Burkina Faso, and (ii) to assess, using additional tests, the biological significance of indeterminate results in order to define a more rational strategy at the individual and public health levels.(This work was presented in part at the

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Section: Discussionmentioning

confidence: 94%

Low Prevalence Rate of Indeterminate Serological Human Immunodeficiency Virus Results among Pregnant Women from Burkina Faso, West Africa

et al. 2010

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“…Since viral load testing requires specialized facilities and experienced personnel, alternative strategies have been explored. Recent reports have suggested that DBS may be suitable for use for the testing of HIV RNA levels, although most studies have so far examined only small sets of samples (1,2,25,32,34). Both the reliability and the sensitivity of tests that use DBS compared to those that use plasma have been matters of concern.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, the use of dried blood spots (DBS) is increasingly being considered as an alternative that can be used to overcome the troubles inherent to the testing of plasma. Patient's blood placed on DBS can be shipped at room temperature to specialized reference laboratories, where the specimens may be subject to HIV RNA quantitation (1,2,3,8,11,22), HIV subtyping, and/or drug resistance testing (5,15,19,28). DBS have also been successfully used to assist with the confirmation of a diagnosis of HIV infection (10,18,27), measurement of CD4 counts (23), and even analysis of antiretroviral drug levels (17).…”

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confidence: 99%

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

Garrido¹,

Zahonero²,

Corral³

et al. 2009

J Clin Microbiol

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The plasma human immunodeficiency virus (HIV) RNA load is used in the clinical routine for the monitoring of HIV infection and the patient's response to antiretroviral therapy. Other body fluids or dried blood spots (DBS) can be used, however, to assess the level of viremia. The use of DBS may be especially helpful for the monitoring of HIV-infected patients in resource-poor settings, where access to adequate laboratory facilities is often difficult. However, the correlation between the HIV RNA levels in plasma and those in DBSs has not been well established. Paired plasma and DBS samples obtained from HIV type 1 (HIV-1)-infected patients were tested for HIV RNA copy numbers by using two different commercial assays, the Nuclisens EasyQ HIV-1 (version 1.1) test (the Nuclisens test; Biomerieux) and the m2000rt RealTime HIV test (the m2000rt test; Abbott). Nucleic acid extraction was performed manually by using either the Nuclisens isolation kit (which uses the Boom methodology) or the m2000rt sample preparation kit (an iron particle-based method). A total of 103 paired plasma and DBS samples were tested. Viral load results were obtained for 97 (94.2%) samples with the Nuclisens isolation kit and 81 (78.6%) samples with the m2000rt kit. The overall correlation between the RNA loads in plasma and DBS was good, although better results were obtained by the Nuclisens test (R 2 ‫؍‬ 0.87, P < 0.001) than by the m2000rt test (R 2 ‫؍‬ 0.70, P < 0.001). While the specificities were excellent and similar for both the Nuclisens and the m2000rt tests (97.1% and 100%, respectively), the sensitivity was greater by the Nuclisens test than by the m2000rt test (75.8% and 56.6%, respectively). Overall, the viral loads in DBS tended to be lower than those in plasma, with mean differences of 0.3 log unit (standard deviation, 0.5 log unit) and 0.76 log unit (standard deviation, 0.8 log unit) for the Nuclisens and the m2000rt tests, respectively. The levels of agreement between the measurements in plasma and DBS were assessed by using the Bland-Altman plot for each assay. The Nuclisens test gave results within its defined limits (؊0.65 to 1.26) for 95.9% of the samples, while the m2000rt test gave results within its limits (؊0.83 to 2.33) for 100% of the samples. In summary, the HIV-1 load can accurately be quantified by testing DBS by either the Nuclisens or the m2000rt test, although the Nuclisens test may outperform the m2000rt test when nucleic acids are extracted manually.

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“…Dried blood spots (DBS) may be an interesting alternative to plasma for periodic VL testing (1,5,7,9,12,23). DBS can be easily collected and stored without being frozen or refrigerated (2,17,22). Several studies have demonstrated the feasibility of DBS as a specimen type for VL testing using different methodologies, although limitations in terms of sensitivity and stability have been noted (3,5,9,10,12,16).…”

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confidence: 99%

Comparison of HIV-1 RNA Measurements Obtained by Using Plasma and Dried Blood Spots in the Automated Abbott Real-Time Viral Load Assay

Arredondo¹,

Garrido²,

Parkin

et al. 2012

J Clin Microbiol

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c Dried blood spots (DBS) may be a promising alternative specimen type to plasma for measuring the viral load (VL) in HIVinfected individuals in resource-limited settings. However, characterization of assay performance using DBS is incomplete. In this prospective study, the VL was measured in parallel using plasma and DBS specimens collected at the same time from 157 HIV-1-infected individuals. DBS were prepared by dispensing 50 l of blood onto filter paper cards and were stored desiccated at ؊20°C. Nucleic acid extraction from plasma and DBS was performed automatically using the Abbott m2000sp instrument, and the VL was measured using the RealTime HIV-1 VL assay, which has a lower limit of detection of 40 HIV RNA copies/ml. The correlation between plasma and DBS results was good (R ‫؍‬ 0.91; P < 0.001). The mean difference in the VL (DBS minus plasma) was 0.35 log copies (standard deviation [SD], 0.47 log copies). A total of 40 (26%) paired specimens had a difference of >0.5 log copy, and in 12 (7.8%) it was >1 log copy. the VL from DBS was measurable in 95.7% of specimens with a plasma VL of >2.74 log copies (550 HIV RNA copies/ml). In summary, the VL can reliably be measured using DBS with the Abbott RealTime HIV-1 assay. The estimated lower limit of detection of this automated methodology on DBS is 550 copies/ml, a threshold that may be acceptable for periodic VL monitoring in patients on antiretroviral therapy in resource-limited settings, where early detection of virologic treatment failure is often problematic. P eriodic measurement of the viral load (VL) in plasma remains a key parameter in the follow-up of HIV-infected individuals, especially those on antiretroviral therapy (ART) (21). However, reliable VL testing in plasma requires specialized facilities that often are not available in resource-poor settings. Dried blood spots (DBS) may be an interesting alternative to plasma for periodic VL testing (1,5,7,9,12,23). DBS can be easily collected and stored without being frozen or refrigerated (2,17,22). Several studies have demonstrated the feasibility of DBS as a specimen type for VL testing using different methodologies, although limitations in terms of sensitivity and stability have been noted (3,5,9,10,12,16). In addition, the recognition of distinct subtypes is an important problem in countries where HIV variability is high, and some VL assays occasionally underquantify some variants (19,24).In a previous study, we reported the results of VL testing on DBS using the Abbott mSample preparation system, a manual RNA isolation method. Although the correlation with plasma values was acceptable, the sensitivity was significantly lower on DBS (ϳ3.5 log copies) (11). Improvements in the detection limit (similar to plasma values of ϳ400 copies/ml) are important for early recognition of virological failure in patients on antiretroviral treatment. Here, we evaluate the feasibility of using an automated RNA isolation method, m2000sp, and the Abbott RealTime HIV-1 VL assay to quantify HIV-1 RNA on DBS and compar...

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Evaluation of the use of dried spots and of different storage conditions of plasma for HIV‐1 RNA quantification

Cited by 35 publications

References 12 publications

Low Prevalence Rate of Indeterminate Serological Human Immunodeficiency Virus Results among Pregnant Women from Burkina Faso, West Africa

Low Prevalence Rate of Indeterminate Serological Human Immunodeficiency Virus Results among Pregnant Women from Burkina Faso, West Africa

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

Comparison of HIV-1 RNA Measurements Obtained by Using Plasma and Dried Blood Spots in the Automated Abbott Real-Time Viral Load Assay

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