ABSTRACT
Objective@#The aim of the present study was to evaluate the performance of the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA using one dried blood spot (DBS) as an alternative sample to plasma.@*Method@#A total of 571 paired DBS/plasma samples were collected from men who have sex with men (MSM) and injection drug users (IDUs), and serological and molecular assays were performed. Using plasma results as the reference standard, the performance of DBS tests for HIV-1 RNA, HIV-1 DNA, and HCV RNA was evaluated. Pearson's correlation coefficients and Bland-Altman analysis were performed to assess the correlation and concordance between DBS and plasma.@*Results@#Among paired plasma/DBS samples with detectable HIV-1 RNA and HCV RNA, five samples (5/32) were not detectable in DBS, while measurable HIV-1 RNA levels were present in plasma (1.44 to 3.99 log @*Conclusion@#The performance of the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA using one DBS was acceptable. DBS, as an alternative sample to plasma, may be a viable option for the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA in resource-limited settings or for individuals living in areas that are difficult to access.
Subject(s)
DNA, Viral/analysis , Diagnostic Tests, Routine/methods , Dried Blood Spot Testing/methods , HIV Infections/diagnosis , HIV-1/isolation & purification , Hepacivirus/isolation & purification , Hepatitis C/diagnosis , RNA, Viral/analysis , Sensitivity and Specificity , Specimen Handling/methods , Syphilis/diagnosis , Treponema pallidum/isolation & purificationABSTRACT
Objectives: To study the correlation between CD4 count &HIV-1 viral load among ART Naive patients attending ICTCSMS Medical College, Jaipur.Material and Methods: This study was conducted on 250 HIVserologically confirmed, ART Naive cases from ICTC, SMSJaipur. RNA extraction was done from plasma samples byQiagen Viral RNA Mini Kit then HIV-1 Viral load wasdetermined by Qiagen HIV-1 viral load kit on ABI 7500 Fast dxReal Time PCR, while CD4 count was done on FACSCALIBUR flowcytometer (BD Biosciences). SPSS ver. 21.0was used to determine correlation between CD4 count & HIV-1viral load.Results: Out of 250, 216 (86.4%) cases were found in whichviral RNA was detected. These samples were correlated withtheir CD4 Count. The mean of viral load was 194746.2791 ±550442.61805 IU/ml while CD4 count was 282.7674 ±217.56456 cells/ul. Females were having Avg. Viral load228506.7273 & CD4 count 337.21 and males were found tohave Avg. Viral load 179791.9866 & CD4 count 258.65Conclusion: This study concluded a negativecorrelation between HIV-1 RNA viral load and CD4 count inHIV-seropositive ART naïve patients of this part of the country.Our study confirmed that HIV-1 RNA viral load levels aresignificantly higher in women than in men, but no suchsignificant gender difference in the CD4 count was found.
ABSTRACT
Purpose: The use of dried blood spots (DBS) for HIV-1 viral load determination could greatly enhance the management of HIV infected individuals in resource-limited countries. Objective: To compare the HIV-1 viral load values obtained between parallel collected plasma and DBS. Materials and Methods: DBS and plasma samples were collected from 62 HIV-1 infected individuals and were used for determination of HIV-1 RNA concentrations using the Abbot real-time HIV-1 PCR. Result: Mean of the log difference of viral load values between plasma and DBS was -0.41 log. DBS viral load values significantly correlated with plasma viral load (r = 0.9818, P < 0.0001). Conclusion: These results suggest that DBS samples can be used as an alternative to plasma for the estimation of HIV-1 viral load if samples are appropriately stored.
Subject(s)
HIV-1/analysis , Humans , India , Patients , Pilot Projects , Real-Time Polymerase Chain Reaction/methods , RNA, Viral/analysis , RNA, Viral/blood , RNA, Viral/isolation & purificationABSTRACT
The question of whether HIV-1 RNA in cerebrospinal fluid (CSF) is derived from viral replication in the central nervous system or simply reflects the transit of infected lymphocytes from the blood compartment has long been a matter of debate. Some studies found no correlation between CSF and plasma viral load, whereas others did. The lack of a correlation between the two compartments suggests that the presence of HIV-1 RNA is not simply due to the passive passage of the virus from blood to CSF but rather due to intrathecal replication. To evaluate the correlation between plasma and CSF HIV-1 RNA levels and to identify situations in which there is no correlation between the two compartments, seventy patients were prospectively studied. The association between CSF and plasma viral load was evaluated in the total population and in subgroups of patients with similar characteristics. A correlation between the CSF and plasma compartments was observed for patients undergoing highly active antiretroviral therapy (HAART), those with a CD4 T lymphocyte count lower than 200 cells/mm³, and those with increased CSF protein content. On the other hand, no correlation was observed for patients without adequate virological control, who had a CD4 count higher than 200 cells/mm³ and who did not use HAART. The correlation between the two compartments observed in some patients suggests that CSF HIV-1 RNA levels may reflect plasma levels in these subjects. In contrast, the lack of a correlation between the two compartments in patients who were not on HAART and who had normal CSF proteins and a poor virological control possibly indicates compartmentalization of the virus in CSF and, consequently, plasma-independent intrathecal viral replication.
Tem sido objeto de debate a questão se o RNA do HIV-1 no líquido cefalorraquidiano (LCR) é derivado da replicação viral no sistema nervoso central ou simplesmente reflete o tráfego de linfócitos infectados do compartimento sanguíneo. Alguns estudos não mostraram correlação entre a carga viral do plasma e LCR, mas outros sim. A falta de correlação entre os dois compartimentos sugere que a presença de RNA do HIV-1 não é simplesmente devido à passagem do vírus do plasma para o LCR, mas sim a uma replicação intratecal. Para avaliar a correlação entre os níveis de RNA do HIV-1 no plasma e no LCR e tentar identificar situações, na qual, não existe a correlação entre os dois compartimentos avaliaram-se setenta pacientes prospectivamente. A associação entre a carga viral do LCR e plasma foi avaliada na população total e em subgrupos de pacientes com características similares. A correlação entre os dois compartimentos foi observada em pacientes que estavam em uso da terapia antiretroviral (HAART), naqueles que tinham contagem de linfócitos CD4 menor que 200 céls/mm³ e naqueles com aumento da concentração de proteínas no LCR. Por outro lado, não houve correlação para os pacientes que não tinham um controle virológico adequado, os que tinham contagem de CD4 maior que 200 céls/mm³ e aqueles que não estavam usando HAART. A correlação entre os dois compartimentos observada em alguns pacientes sugere que os níveis de RNA do HIV-1 no LCR podem refletir os níveis plasmáticos nestes pacientes. E a falta de correlação ente os dois compartimentos em pacientes que não usavam HAART, nos que tinham uma concentração de proteínas no LCR normal, e nos que não apresentavam bom controle virológico, indica provavelmente a compartimentalização do vírus no LCR e consequentemente replicação viral intratecal independente da do plasma.