BCN Checkpoint: same-day confirmation of reactive HIV rapid test with Point Of Care HIV-RNA accelerates linkage to care and reduces anxiety.

BACKGROUND: The introduction in 2006 of the rapid HIV test by BCN Checkpoint in a non-clinical setting has been a successful step forwards in the uptake of testing. Nevertheless, HIV serostatus should be reported as HIV positive only when a reactive result has been tested again using a different assay (WHO guidelines 2015). The standard confirmation test has been the Western Blot (WB) test. However confirmation results take around 7 days to come back. AIMS: This study explores the possibility of Point of Care PCR testing for a same-day confirmation. MATERIALS AND METHODS: Between March 2015 and September 2016 a POC PCR test (Xpert® HIV-1 Qual) was performed in parallel to the Western Blot test after a reactive HIV rapid test (Alere Determine™ HIV-1/2 Ag/Ab Combo and Alere™ HIV Combo). HIV confirmed positive cases received emotional support by peers, were informed and prepared for treatment initiation and rapidly linked to HIV clinic. RESULTS: During the study period 11 455 tests were performed to 7163 clients. A total of 249 reactive rapid HIV tests were found. For analysis a total of 33 cases were excluded due to the lack of PCR and/or WB test. Results of comparison of the 216 cases showed 194 concordant positive confirmations and 14 concordant negative results. In three cases PCR was positive and WB negative. In five cases PCR was negative and WB positive. CONCLUSION: The POC PCR assay is easy to use and feasible in a community-based center. Reducing time for confirmation to 90 min has been possible in 91.2% (197/216) of cases with positive PCR result. In cases of a negative PCR result an additional test (WB, Elisa or PCR quantitative) was needed to distinguish false positive results (6.5%) from viral load results below level of detection (2.3%). Clients expressed satisfaction with same-day confirmation and less anxiety.

[Evaluation of immunity against Venezuelan equine encephalitis virus and dengue in the human population of San Carlos, the Almirante Padilla Island Municipality, Zulia State, Venezuela. 1996]. / Evaluación de la inmunidad contra los virus de encefalitis equina venezolana y dengue en la población humana de San Carlos, Municipio Insular Almirante Padilla, Estado Zulia, Venezuela. Año 1996.

The Insular Municipality of Almirante Padilla, historically, has been affected by Venezuelan Equine Encephalitis (VEE) epidemics, that have occurred cyclically in the Paez, Mara and Almirante Padilla Municipalities of the State of Zulia. During the last epidemic outbreak (1995), the studies reported a great number of cases of VEE in humans, based on epidemiologic-clinical diagnosis; occupying this municipality, the first place in the attack rate by the virus. At the same time, Dengue has been defined as an endemic illness affecting different regions, whose etiologic agent has previously circulated in the studied zone. In order to evaluate the immunity acquired against these viruses, a serological study was conducted in San Carlos Island. Two hundred and ten blood samples were obtained at random, among individuals of an age range between < 1 and 69 years, with and without antecedents of viral illness symptomatology at the moment of the epidemic outbreak, occurred in 1995. The samples were classified according to sex and age, and analyzed through the test of ELISA for specific IgG antibodies against the VEE and Dengue viruses. From the total of the samples processed, 116 (55.2%) were positive for VEE, affecting uniformly all age-groups with a slight masculine predominance. Likewise, 88 cases (41.9%) were determined positive for Dengue virus, affecting mainly the group ranging from 10 to 19 years. These results let us suggest that the epidemic outbreak occurred in 1995 at the Almirante Padilla Insular Municipality, had a double nature with both viruses being involved in the etiology of febrile cases.

Melatonin treatment enhances the efficiency of mice immunization with Venezuelan equine encephalomyelitis virus TC-83.

To determine whether treatment with melatonin (MLT) improves the efficiency of immunization against Venezuelan equine encephalomyelitis (VEE) virus, mice were vaccinated with TC-83 VEE virus and treated daily with MLT (1 or 5 mg/kg) starting 3 days before immunization, until 10 days after. IgM antibody titers were determined at days 7, 14, and 21 post-immunization. IL-10 levels were assayed at day 14 postvaccination. Treatment with MLT increased antibody titers 14 days after the immunization. IL-10 levels also increased with MLT treatment (1 and 5 mg/kg). Mice were challenged with live VEE virus at day 21 postimmunization, and viral titers were plaque assayed in chicken embryo fibroblasts 4 days after the infection. Following this challenge brain virus levels were significantly reduced. The results suggest that MLT treatment enhances the efficiency of mice immunization against VEE virus.

High intensity light increases olfactory bulb melatonin in Venezuelan equine encephalitis virus infection.

In mice infected with the Venezuelan equine encephalomyelitis (VEE) virus and exposed to high intensity light (2500 lux) with a 12 h light: 12 h dark photoperiod, a significant increase in the levels of melatonin in the olfactory bulb was observed. The significance of these findings deserves further studies to understand the mechanisms involved in this effect since the olfactory bulbs have been proposed as first portal for VEE virus entry into the CNS. The increase in melatonin content could represent one of the mechanisms of defense against the viral attack.

Melatonin prolongs survival of immunodepressed mice infected with the Venezuelan equine encephalomyelitis virus.

Male albino mice immunodepressed after the injection of dexamethasone (DEX) were inoculated intraperitoneally with the Guajira strain of Venezuelan equine encephalomyelitis (VEE) virus. Melatonin (MLT) was administered daily, at a dose of 500 micrograms/kg bodyweight, for 3 days before virus inoculation and 10 days after. Serum levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-2 (IL-2) were determined in all the experimental groups (control, DEX, DEX + MLT, DEX + VEE, DEX + VEE + MLT, VEE and MLT). At day 6 after the virus inoculation, the survival rate was significantly increased from 0% in group DEX + VEE to 32.5% in the group of immunodepressed infected mice treated with MLT (DEX + VEE + MLT). By day 10 a survival rate of 10% was found in group DEX + VEE + MLT and 0% in group VEE. No alterations in IL-2 serum levels were observed. MLT increased GM-CSF in control and in DEX-treated mice. In the VEE virus-infected mice treated with DEX, serum levels of GM-CSF increased progressively from day 1 to 5 postinoculation. In contrast, the levels of GM-CSF in infected immunodepressed mice treated with MLT decreased significantly from day 1 to 5 postinoculation. At day 5 after viral inoculation, no differences were detected in the cerebral viral titres in groups VEE, DEX + VEE and DEX + MLT + VEE. These results show that MLT does not inhibit VEE viral replication in the brain of immunodepressed mice.

In mice the efficiency of immunization with Venezuelan Equine Encephalomyelitis virus TC-83 is transiently increased by dehydroepiandrosterone.

To determine whether treatment with dehydroepiandrosterone (DHEA) improves the efficiency of immunization against the Venezuelan Equine Encephalomyelitis (VEE) virus, mice were vaccinated with the TC-83 VEE virus. DHEA (10 mg/kg) was administered in a single dose, 4 hours before vaccination. IgM antibody titers were determined at days 7, 14 and 21 post-immunization. Treatment with DHEA increased antibody titers at day 14 after immunization. Mice were challenged with live VEE virus at day 21, and viral titers were plaque assayed in chicken embryo fibroblasts from days 2 to 5 post-infection. After the challenge, viremia decreased on day 2 and brain virus levels were reduced at day 4 in mice treated with DHEA. These results suggest that DHEA treatment could enhance the efficiency of immunization against VEE virus in mice.

Exposure to 2500 lux increases serum melatonin in Venezuelan equine encephalomyelitis.

When mice infected with the Venezuelan equine encephalomyelitis (VEE) virus were exposed to 2500 lux with a 12 h light: 12 h dark photoperiod, the serum levels of melatonin (MLT) remained constantly elevated. In mice exposed to 400 lux low levels of serum MLT were detected during the day and high levels during the night. An increase in the survival rate of the infected mice from 6 to 13 days after virus inoculation was also observed. The significant increment in the concentration of serum MLT produced by the high intensity light could be responsible for the longer survival rate of mice infected with the VEE virus.

[Effect of the attenuated strain (TC-83) of Venezuelan equine encephalitis virus on nuclear transcription in rat brain cells]. / Efecto de la cepa atenuada (TC-83) del virus de la Encefalitis Equina Venezolana sobre la transcripción nuclear en las celulas cerebrales de ratas.

In the present study the effect of the attenuated strain TC-83 of the Venezuelan Equine Encephalitis virus on the nuclear transcription in brain cells of rats was assessed. The transcription activity of the DNA depending RNA polymerases (types I and II) in the isolated nuclei of brain of infected rats and controls was determinated by incorporation of the (3H) UTP. Simultaneously a viral replication curve in the brain and the serum was carried out by plaque forming method in chicken embryo cell cultures. RNA polymerase I activity was only significantly reduced after 25 hours of infection, respect to control values, while polymerase II activity was progressive and significantly diminished from inicial stages of the viral infection at 10, 15, 20 y 25 hours post-infection compared to control values. The virus was not detected in the brain but after 25 hours post-infection with very low titers (< 0.7 log10 P.F.U./ml.), while the viral presence in the blood was demonstrated after a 10 hour period. Our results demonstrated a marked effect of the attenuated strain on the brain nuclear transcription, although the presence of the virus was not detected in the brain of the infected rats. This finding suggest a mechanism of action which deserves further studies to elucidate the cerebral metabolic response and the pathogenesis of the Venezuelan Equine Encephalitis infection.

[Venezuelan equine encephalitis. Determination of antibodies in the human population of Municipio Mirand, Estado Zulia, Venezuela]. / Encefalitis equina venezolana. Determinación de anticuerpos en la población humana del Municipio Miranda, Estado Zulia, Venezuela.

With the purpose of determining antibodies prevalence against Venezuelan Equine Encephalitis Virus in the population of Puertos de Altagracia and Sabaneta de Palmas of Miranda county, Zulia State, Venezuela, 199 subjects were studied: 57 from Puertos de Altagracia and 142 from Sabaneta de Palmas. They were classified in older (42.78%) and younger (57.2%) than 15 years. The blood specimens were processed for Hemagglutination Inhibition Test using EEV antigen Goajira strain at pH 6.5. We found that all 57 specimens from Puertos de Altagracia were negative, whereas of 142 specimens from Sabaneta de Palmas 17 were positive (11.97%). Of these, one was from a subject less than 15 years-old (5.85%) and 16 from individuals more than 15 years-old (94.15%). Positive titers were higher than 1:160 in 80% of cases. Being Sabaneta de Palmas one of the most affected areas in the 1962 epidemic in the Miranda county and keeping the affected ones high positivity with elevated titers, we conclude that this population could represent an enzootic zone similar to Paez county where a similar situation, of high positivity and elevated titers, many years after the last epidemic occurred in that area, has been described.