Dried blood spots versus plasma for the quantitation of HIV-1 RNA using a real-Time PCR, m2000rt assay

Madhavan, Vidya; Saravanan, S.; Rifkin, Samara; Solomon, Sunil S.; Waldrop, Greer; Mayer, Kenneth H.; Solomon, Suniti; Balakrishnan, Pachamuthu

doi:10.1016/j.jviromet.2012.02.006

Cited by 31 publications

(36 citation statements)

References 31 publications

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“…However, an overquantification of the VL in DBS was observed in pairs with a plasma VL of Ͻ3,000 copies/ml, likely due to proviral DNA and intracellular RNA contaminations. Similar observations were recently made by Vidya and colleagues (23) and they reported the same threshold of 3,000 copies/ml. Other reports have also mentioned the reduced performance of the m2000rt assay on DBS for a plasma VL of Ͻ3,000 copies/ml (18,24).…”

Section: Discussionsupporting

confidence: 89%

Field Evaluation of Dried Blood Spots for Routine HIV-1 Viral Load and Drug Resistance Monitoring in Patients Receiving Antiretroviral Therapy in Africa and Asia

et al. 2014

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h Dried blood spots (DBS) can be used in developing countries to alleviate the logistic constraints of using blood plasma specimens for viral load (VL) and HIV drug resistance (HIVDR) testing, but they should be assessed under field conditions. Between 2009 and 2011, we collected paired plasma-DBS samples from treatment-experienced HIV-1-infected adults in Burkina Faso, Cameroon, Senegal, Togo, Thailand, and Vietnam. The DBS were stored at an ambient temperature for 2 to 4 weeks and subsequently at ؊20°C before testing. VL testing was performed on the plasma samples and DBS using locally available methods: the Abbott m2000rt HIV-1 test, generic G2 real-time PCR, or the NucliSENS EasyQ version 1.2 test. In the case of virological failure (VF), i.e., a plasma VL of >1,000 copies/ml, HIVDR genotyping was performed on paired plasma-DBS samples. Overall, we compared 382 plasma-DBS sample pairs for DBS VL testing accuracy. The sensitivities of the different assays in different laboratories for detecting VF using DBS varied from 75% to 100% for the m2000rt test in labs B, C, and D, 91% to 93% for generic G2 realtime PCR in labs A and F, and 85% for the NucliSENS test in lab E. The specificities varied from 82% to 97% for the m2000rt and NucliSENS tests and reached only 60% for the generic G2 test. The NucliSENS test showed good agreement between plasma and DBS VL but underestimated the DBS VL. The lowest agreement was observed for the generic G2 test. Genotyping was successful for 96/124 (77%) DBS tested, and 75/96 (78%) plasma-DBS pairs had identical HIVDR mutations. Significant discrepancies in resistance interpretations were observed in 9 cases, 6 of which were from the same laboratory. DBS can be successfully used as an alternative to blood plasma samples for routine VL and HIVDR monitoring in African and Asian settings. However, the selection of an adequate VL measurement method and the definition of the VF threshold should be considered, and laboratory performance should be monitored.T he increasing availability of antiretroviral treatment (ART) has dramatically contributed to a reduction in mortality and morbidity related to HIV/AIDS in resource-limited countries (RLC). In order to limit the emergence of resistance to antiretroviral drugs, HIV treatment should ideally be accompanied by regular virological monitoring, including viral load (VL) and genotypic drug resistance testing (1). With the recent scaling up of ART in RLC, people from semiurban and rural areas are now also receiving treatment, and recent data have shown variable levels of virological failure (VF) and drug resistance in these areas. In certain settings, VF can be Ͼ20% after 12 months of ART (2-4), stressing the urgent need for improved virological monitoring.In the majority of developing countries with limited resources, monitoring blood plasma VL poses logistical challenges since plasma preparation, storage, and/or shipment requires personnel and laboratory infrastructure that is often lacking. Today, viral load assays still require sophistic...

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Section: Discussionsupporting

confidence: 89%

Field Evaluation of Dried Blood Spots for Routine HIV-1 Viral Load and Drug Resistance Monitoring in Patients Receiving Antiretroviral Therapy in Africa and Asia

et al. 2014

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“…[5] The modest overquantification that we observed with DBS can be ascribed to the presence of HIVDNA and intracellular RNA and seems to have a greater impact on samples with a low viral load (in our study <3 000 copies/mL), as previously reported. [4,11,14] The 2013 HIV treatment guidelines of the World Health Organi zation [15] indicate a threshold of 1 000 copies/mL of HIVRNA to define virological failure using plasma, and suggest use of a higher, although undefined, threshold when using DBS. In previous studies performed with the Abbott system, sensitivity for the detection of virological failure at 1 000 copies/mL was between 75% and 100% and specificity was between 82% and 97%.…”

Section: Discussionmentioning

confidence: 99%

“…Nucleic acids in DBS have been shown to be stable for several months at room temperature, provided the DBS specimens are thoroughly dried and are stored with desiccant. [14] DBS specimens can therefore be collected at remote rural sites and transported to a central or regional testing laboratory without refrigeration. DBS are userfriendly and costeffective in resourcelimited settings, their ability being demon strated to correctly identify virological failure.…”

mentioning

confidence: 99%

Measurement of viral load by the automated Abbott real-time HIV-1 assay using dried blood spots collected and processed in Malawi and Mozambique

Erba

Brambilla²,

Ceffa³

et al. 2015

S Afr Med J

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Background. The use of dried blood spots (DBS) for HIV1 viral load quantification can greatly improve access to viral monitoring for HIVinfected patients receiving treatment in resourcelimited settings. Objectives. To evaluate and validate HIV viral load measurement from DBS in subSaharan Africa, with a reliable, allautomated, standard commercial assay such as the Abbott m2000. Methods. A total of 277 DBS were collected in different health centres in Malawi and Mozambique and analysed for viral load determination using the Abbott m2000 assay with the corresponding plasma samples as gold standard. Samples were extracted using the m2000SP automatic extractor and then processed as the plasma samples using the specific 1.0 mL HIVRNA DBS protocol. Results. Among samples with detectable HIVRNA the correlation between viral load obtained from the paired 131 plasma and DBS samples was high (r=0.946). Overall, viral load values between DBS and plasma differed by less than 0.5 log unit in 90.1% of cases and by less than 1 log unit in 100% of cases. Using a threshold of 1 000 copies/mL (defining virological failure in resourcelimited settings), sensitivity was 94.2% and specificity 98.6%, and both positive and negative predictive values were high (98.5% and 94.5%, respectively). Conclusion. DBS extracted and processed using the Abbott automated system can be reliably used in resourcelimited setting to diagnose virological failure.

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“…In some resource-limited countries, the use of dried blood spots (DBS) for sample collection has been proven to be able to keep the viral nucleic acid in good condition during transportation. The cost of using filter paper for DBS sampling is much more cost-effective than using plasma preparation tubes or ethylenediaminetetraacetic acid tubes for whole-blood collection [25]. Using the Abbott HIV-1 Realtime assay (Abbott Molecular, USA), the RNA quantitative levels were not significantly different between freshly separated plasma and DBS.…”

Section: Viral Load Monitoringmentioning

confidence: 99%

Current Assays for HIV-1 Diagnostics and Antiretroviral Therapy Monitoring: Challenges and Possibilities

Chen

Yam

2013

Future Virol.

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In 2011, there were over 34 million people living with HIV infections, placing a heavy burden on public health sectors. HIV infection is a lifelong threat that cannot be prevented by vaccination or cured by antiretroviral drugs. The infected patients rely on daily antiretroviral therapy to suppress HIV viral replication. Hence, it is important to diagnose HIV infections as early as possible and to monitor the efficacy of antiretroviral therapy every 3–6 months. Different immunoassays detecting HIV antigens and antibodies have been modified to offer better sensitivity and more rapid diagnosis. Several clinical and virological parameters, including CD4+ cell counts, viral load and drug resistance mutations, are also used for treatment monitoring. Many molecular assay optimizations are now being utilized to improve patient care. This review will focus on the most updated HIV diagnostic assays, as well as discussing the upcoming possibilities of other advanced technologies.

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Dried blood spots versus plasma for the quantitation of HIV-1 RNA using a real-Time PCR, m2000rt assay

Cited by 31 publications

References 31 publications

Field Evaluation of Dried Blood Spots for Routine HIV-1 Viral Load and Drug Resistance Monitoring in Patients Receiving Antiretroviral Therapy in Africa and Asia

Field Evaluation of Dried Blood Spots for Routine HIV-1 Viral Load and Drug Resistance Monitoring in Patients Receiving Antiretroviral Therapy in Africa and Asia

Measurement of viral load by the automated Abbott real-time HIV-1 assay using dried blood spots collected and processed in Malawi and Mozambique

Current Assays for HIV-1 Diagnostics and Antiretroviral Therapy Monitoring: Challenges and Possibilities

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