Influenza B false-positive results by rapid molecular tests Alere™ i Influenza A&B 2 in France.

The Alere-i™ Influenza A&B (Abbott), a nicking endonuclease amplification reaction test, has recently been improved in order to deliver results in a few minutes. Our field observation highlights two problems with this new version: improper interpretation of a test as valid despite improper reagent hydration and falsely influenza B positive results. We advise users of the new system to check reagent hydration prior to reporting a result and to systematically confirm positive influenza B results.

Failure of combined antiretroviral therapy intensification with maraviroc and raltegravir in chronically HIV-1 infected patients to reduce the viral reservoir: the IntensHIV randomized trial.

BACKGROUND: Ongoing HIV-1 replication in lymphoid cells is one explanation of the persistence of HIV-1 reservoirs despite highly active antiretroviral therapy (cART). We tested the potential of cART intensification by Maraviroc plus Raltegravir to decrease proviral HIV-1 DNA levels in lymphoid cells during a randomized trial. PATIENTS AND METHODS: We randomly assigned for 48 weeks 22 patients to continue their current first line regimen of Truvada® plus Kaletra® or intensify it with Maraviroc and Raltegravir. The primary objective was to obtain a 50% decrease in proviral HIV-1 DNA levels in lymphoid cells with intensification. Blood samples were drawn at W-2, W0, W2, W4, W12, W24 and W48. Plasma viremia, cellular proviral DNA and cellular RNA, 2-LTR circles and lymphocytes subsets were assayed using validated methods. Patients in the intensified group underwent a gut biopsy at baseline and W48 to measure proviral DNA levels. Statistical analysis used parametric and non-parametric tests. RESULTS: Ten patients in each arm completed the trial. The 2 populations were comparable at baseline. No change in the reservoir size was observed in the intensified arm compared to the control arm measured in peripheral blood mononuclear cells (PBMCs). No change in the reservoir size was observed in gut proviral DNA in the intensified arm. In this group, no increase in 2-LTR circles was observed as early as 2 weeks after intensification and no change was found in residual plasma RNA levels measured by the single copy assay. However, a decrease in CD8(+) T cells activation was observed at 24 and 48 weeks, as well as in PBMCs HIV-1 RNA levels. CONCLUSION: We conclude that the intensification of a Protease Inhibitor regimen with Maraviroc and Raltegravir does not impact the blood proviral DNA reservoir of HIV but can decrease the cell-associated HIV RNA, the CD8 activation and has a possible impact on rectal proviral HIV DNA in some patients. TRIAL REGISTRATION: ClinicalTrials.gov identifier number NCT00935480.

Hepatitis E virus in HIV-infected patients.

OBJECTIVES: Many cases of acute autochthonous hepatitic E virus (HEV) hepatitis have been reported in France, mainly from the south. Chronic HEV infection has recently been described in immunosuppressed patients. Although a potential risk of chronicity exists in HIV-infected patients, no survey has been conducted in this population. The aim of this study was to assess the sero-virological prevalence of HEV in French HIV-infected patients. METHODS: Two hundred and forty-five HIV-infected patients followed at two Infectious Diseases Departments (one in the south, one in the north) were included from January to March 2009. Sera were collected from all patients and tested using anti-HEV IgG and IgM kits. HEV RNA was systematically amplified in the ORF2 region with an in-house method. The IgG avidity index of all IgG-positive samples was determined. RESULTS: Three of the 133 southern patients showed both anti-HEV IgG and IgM positivities, along with cytolysis and biological cholestasis; HEV RNA was amplified in two of these cases, whereas a low IgG avidity index was observed in all three samples. Twelve of the 130 remaining southern patients (9%) showed anti-HEV IgG positivity. The serological prevalence in the 112 northern patients was 3%, which was significantly lower than in the southern patients (P=0.04). No case of acute hepatitis was reported in the north, whereas the prevalence of patients with biochemical liver abnormalities was similar in both areas (P=0.22). CONCLUSIONS: In France, HIV-infected patients are at risk of HEV infection with a serological north-to-south gradient. No case of chronic HEV infection was detected in this study.

Rectal cell-associated HIV-1 RNA: a new marker ready for the clinic.

Gut-associated lymphoid tissue is a huge reservoir for HIV-1. Developing new strategies to target "residual" HIV-1 in patients on effective therapy brings the need for an evaluation of tissue reservoirs in the clinic. We measured cell-associated HIV-1 RNA and DNA in blood and rectal biopsies from 23 patients, including 14 with undetectable viremia on HAART, by using an adaptation of commercially available tests. Rectal cell HIV-1 RNA was detected in all viremic patients, with median levels of 4.90 log(10) copies/million CD4. Although plasma viremia was found at a median of 3 copies/mL in "aviremic" patients, rectal cell HIV-1 RNA was detected in 28.5% with median levels of 5.17 log(10) copies/million CD4. Consequently, we propose to use this marker in future clinical trials targeting "residual" HIV-1 in patients with viremia below the detection limit.

Prevalence of complete resistance to at least two classes of antiretroviral drugs in treated HIV-1-infected patients: a French nationwide study.

The objective of the study was to estimate the prevalence of HIV-1 resistance to all drugs belonging to two or more antiretroviral drug (ARV) classes in treated patients in France. All genotyping assays performed in June 2001 and in November 2002 by the ANRS resistance laboratory network were analyzed by the ANRS algorithm. The 17 and 21 centers of the ANRS network participating in the study in 2001 and 2002, respectively, genotyped the viruses in plasma of 456 and 529 patients, respectively. In 2002, the proportions of patients harboring viruses fully resistant to one, two, and three ARV classes were 5.1%, 8.1%, and 2.5%, respectively. These results were similar to those obtained in 2001. In 2002, among the 56 patients with viruses completely resistant to at least two ARV classes, 98%, 96%, and 29% of patients had viruses with complete class resistance to NRTIs, NNRTIs, and PIs, respectively. Complete resistance to PIs was less frequent than full resistance to the other two ARV classes, and ritonavir-boosted amprenavir and lopinavir/r remained potentially active in respectively 71.4% and 42.9% of these 56 patients. In 2001 and 2002, respectively 30% of the 65 patients and 24% of the 56 patients with viruses completely resistant to at least two ARV classes were at an advanced stage of HIV disease, with CD4(+) cell counts below 200/microl and viral loads above 30 000 copies/ml. In France, the prevalence of HIV-1 viruses completely resistant to two or more ARV classes remained stable between 2001 and 2002. Resistance to RT inhibitors was more frequent than resistance to PIs in patients with viruses completely resistant to two or three classes of ARV.

French national sentinel survey of antiretroviral drug resistance in patients with HIV-1 primary infection and in antiretroviral-naive chronically infected patients in 2001-2002.

OBJECTIVE: To survey the frequency of genotypic antiretroviral resistance and the spread of non-B subtypes in patients with primary HIV-1 infection (2001-2002) and in treatment-naive chronically HIV-1-infected patients (2001). METHODS: Plasma samples from 303 patients with acute HIV-1 infection (Primo study) and 363 treatment-naive patients with chronic HIV-1 infection (Odyssee study) were tested for genotypic resistance. Resistance mutations were identified from the International AIDS Society Resistance Testing-USA panel and resistant viruses were defined according to the French Agence Nationale de Recherches sur le SIDA (ANRS) resistance algorithm. RESULTS: In the Primo study, 14% of the patients had viruses with resistance mutations and 12% of patients had viruses with mutations conferring resistance to least 1 antiretroviral drug. Thirty patients had viruses with mutations to at least 1 antiretroviral drug in a single pharmacologic class. Six patients were infected by viruses resistant to 2 or 3 classes of drugs. In the Odyssee study, the prevalence of reverse transcript (RT) associated and major protease inhibitor-associated mutations was 6.1% (95% CI: 3.6-8.6). Six patients had viruses resistant to at least 1 antiretroviral drug and 3 patients had viruses resistant to 2 classes of antiretroviral drugs. Twenty-four percent of acutely infected patients harbored non-B subtype strains (19% in 1999-2000) and 33.2% of chronically infected patients (10% in 1998; P < 0.0001). CONCLUSION: In France, the frequency of HIV-1 resistance in untreated patients was not significantly higher in 2001-2002 than in previous surveys while the prevalence of non-B subtypes is increasing.

Metabolic evaluation of HIV-infected patients receiving a regimen containing lopinavir/ritonavir (Kaletra).

PURPOSE: We have analyzed retrospectively the evolution of metabolic parameters in a cohort of 159 HIV-infected patients taking a lopinavir/ritonavir-containing regimen during a mean period of 15 months. METHOD: This study was completed by an additional evaluation after strict 12 hours fasting of total cholesterol (TC), HDL-c, LDL-c, triglycerides (TG), glucose, and insulin levels in a subset of 100 patients from the cohort. RESULTS: TC and TG levels increased early after introduction of lopinavir/ritonavir, but remained subsequently stable. After a median of 15 months, TC was greater than normal laboratory range in 46% of cases and TG levels were greater than normal laboratory range in 52% of cases. However, in nearly 90% of these cases, elevations were less than or equal to grade 2. These increases were dependant on CDC stage and lipid levels at lopinavir/ritonavir initiation. No impact on glucose metabolism was found. CONCLUSION: These results are particularly reassuring for the use of lopinavir/ritonavir in everyday practice.

The HYDILE trial: efficacy and tolerance of a quadruple combination of reverse transcriptase inhibitors versus the same regimen plus hydroxyurea or hydroxyurea and interleukin-2 in HIV-infected patients failing protease inhibitor-based combinations.

PURPOSE: To compare the efficacy and tolerance of a stavudine (d4T), didanosine (ddI), efavirenz (EFV), and abacavir (ABC) combination regimen with an identical regimen plus hydroxyurea (HU), or plus HU and interleukin-2 (IL-2), in patients failing protease inhibitor-based combinations and naive of EFV and ABC. METHOD: This was a randomized prospective trial in 69 HIV-infected patients recruited in one clinical center. Antiretroviral drugs were administered at standard doses according to weight. HU was added at week 6 at 500 mg twice daily. Three courses of IL-2 were given subcutaneously at 4.5 MU twice daily for 5 consecutive days, between weeks 24 and 40. The proportion of patients reaching plasma HIV-1 RNA <200 and <50 copies/mL was compared in the three trial groups at weeks 6, 24, and 48 using intent-to-treat and as-treated analyses. CD4+ T-cell count changes from baseline were also assessed at the same time points, along with anthropometric and metabolic measurements. RESULTS: After 48 weeks, only 25% of patients receiving antiretrovirals had plasma HIV-1 RNA <200 copies/mL versus 59.1% in the group receiving HU and 56.5% in the group receiving HU and IL-2 (intent-to-treat; p <.01). At the 50 copies/mL cutoff, the results were 20.8%, 54.5%, and 47.8%, respectively. Most treatment discontinuations were due to failure in the first group and adverse events in the two others. A median decline of 27 CD4+ cells was observed in patients receiving antiretrovirals plus HU, against a gain of 78-118 cells at week 48 in patients receiving antiretrovirals alone or in combination with HU and IL-2. More patients were affected by clinical fat atrophy symptoms at week 48 than at baseline. Additionally, a trend toward increased cholesterol levels was observed throughout the study. CONCLUSION: During this trial, virologic response in patients failing previous regimens was clearly enhanced by the addition of HU, despite d4T and ddI recycling. Although adverse events were more frequent in the HU-containing arms, no unexpected toxicity was observed and the blunted CD4 response prompted by HU was corrected by the addition of IL-2. The combination of HU with reverse transcriptase inhibitors can therefore be regarded as a valuable alternative for patients with few remaining therapeutic options.