Effect of Antiviral Therapy on the Outcome of Mechanically Ventilated Patients With Herpes Simplex Virus Type 1 in BAL Fluid: A Retrospective Cohort Study.

BACKGROUND: Herpes simplex virus type 1 (HSV-1) is frequently detected in the BAL fluid of patients on mechanical ventilation. RESEARCH QUESTION: The aim of the study was to investigate whether antiviral therapy is associated with improved overall survival within 30 days. STUDY DESIGN AND METHODS: This was a retrospective cohort study in four ICUs between January 2011 and December 2017. All adult patients on mechanical ventilation with a respiratory tract infection with positive polymerase chain reaction testing for HSV-1 in the BAL were included. Patients already receiving antiviral agents on the day BAL was performed were excluded. We performed uni- and multivariable Cox and logistic regression modeling. RESULTS: Overall, 306 patients were included in the analysis. Among them, 177 patients (57.8%) received antiviral therapy (90.9% acyclovir, 6.2% ganciclovir, 2.9% both). The overall 30-day mortality rate was 42.4% (n = 75) in the antiviral treatment group and 50.4% (n = 65) in the control group. The adjusted hazard ratio (HR) for the primary outcome was 0.62 (95% CI, 0.44-0.87; P = .005), indicating better overall survival within 30 days for the antiviral-treated group than for the untreated group. This benefit was also present in the subgroup of patients without immunosuppression (n = 246; adjusted HR, 0.53; 95% CI, 0.36-0.78; P = .001). Overall, the median lengths of hospital stay (31 vs 24 days, P = .002) and ICU stay (24 vs 17 days, P < .001), and the duration of mechanical ventilation (18 vs 11 days, P < .001), were longer for patients with therapy. No evidence for the treatment-related deterioration of renal function was observed. INTERPRETATION: These data suggest that detection of HSV-1 in the BAL of patients on mechanical ventilation may be of clinical significance and that specific antiviral treatment may improve clinical outcomes. However, this needs to be proven in multicenter randomized controlled trials before implementation into the clinical routine.

A retrospective analysis of the application of the Elecsys® HIV combi PT assay in southern China.

BACKGROUND: Fourth-generation HIV assays have been implemented worldwide as a screening test for many years. Understanding the performance of fourth-generation assay in low HIV prevalence region is pivotal to interpret the test result correctly. In this study, retrospective analysis was used to evaluate application of the Elecsys® HIV combi PT assay. METHODS: A total of 85 043 specimens from a low prevalence setting were detected between June 2013 and October 2015. We evaluated the false-positive rate (FPR), specificity, and positive predictive value (PPV). RESULTS: The specificity between male and female were 99.85% and 99.82%, respectively. The PPV on male (50.75%) was higher than female (17.05%) significantly, while the FPR was 0.15% and 0.18%. The gap between false-positive (median: 1.83, [IQR]: 1.30, 3.38) and confirmed-positive (median: 407.5, [IQR]: 184.2, 871.7) is enormous. The highest s/co ratio for false-positive cases was 85.45, while the lowest s/co ratio for confirmed-positive cases was 59.68. Various reasons were attributed to false-positive cases. CONCLUSION: Optimal cutoff value is needed to be set to reduce the false-positive cases and predict the final status of HIV infection reliably. Retrospective analysis will help us to understand more about diagnosis of HIV.

Can Analysis of Routine Viral Testing Provide Accurate Estimates of Respiratory Syncytial Virus Disease Burden in Adults?

Respiratory syncytial virus (RSV) is increasingly recognized as a significant cause of adult respiratory illness. We evaluated routine viral testing and discharge diagnoses for identifying RSV and influenza burden. Polymerase chain reaction results performed in adults during emergency room visits or hospitalizations were reviewed. Peak RSV activity preceded influenza activity by 8 weeks. The ratio of total number of viral tests performed divided by total number of respiratory visits was higher during influenza than RSV peaks (1.31 vs 0.72; P = .0001). Influenza and RSV were listed primary diagnoses in 56 (30%) vs 7 (6%), respectively (P < .0001). Routine viral testing to estimate adult RSV disease burden has limitations.

Seguimiento del primer año de tratamiento antirretroviral en pacientes naive en un hospital de tercer nivel / First year monitoring of antiretroviral therapy in naïve patients in a tertiary hospital

Objetivo: El objetivo principal de este trabajo es determinar la tasa de fracaso virológico (FV) en pacientes naive que inician tratamiento antirretroviral (TAR) a 24 y 48 semanas en vida real en un hospital de tercer nivel. Material y método: Estudio retrospectivo de 3 años de duración. Se seleccionaron pacientes adultos VIH naive que iniciaron TAR entre 2012 y 2014. Se registraron datos demográficos (edad, sexo y nacionalidad), clínicos (mecanismo de trasmisión y estadio clínico), de laboratorio (carga viral (CV), linfocitos CD4 basales y existencia de test de mutaciones previo) y TAR elegido. Tras 24 y 48 semanas se registraron: CV y CD4, adherencia, seguimiento, problemas relacionados con la medicación, cambios de TAR y motivos de cambio. Resultados: Se seleccionaron 253 pacientes. 244 y 226 contaban con datos analíticos a las 24 y 48 semanas respectivamente. 142 (58,23%) tenían CV<50 copias/ml y 204 (83,6%) CV<200 copias/ml tras 24 semanas. 198 (88,4%) tenían CV<50 copias/ml y 217 (96,0%) CV<200 copias/ml tras 48. La CV basal superior a 100.000 copias/ml y la mala adherencia se asociaron con un mayor riesgo de FV. Al 30,0% de pacientes se les cambió el TAR durante el primer año, principalmente por efectos secundarios (31,6%) y simplificación del tratamiento (23,7%). Discusión y Conclusiones: Un alto porcentaje de pacientes que inician TAR están indetectables o con menos de 200 copias/ml dentro del primer año de tratamiento. A pesar de tener fármacos altamente eficaces y cada vez mejor tolerados, los efectos secundarios siguen siendo el motivo mayoritario de cambio de TAR.

Background and objective: The main objective of this work is to determine the rate of virologic failure (VF) intreatment-naïve patients, 24 and 48 weeks after starting antiretroviral therapy (ART) in real life in a tertiary hospital. Patients and Methods: A retrospective study of 3 years duration. Naïve HIV adult patients who started ART between 2012 and 2014 were selected. Demographics (age, sex and nationality), clinical (transmission mechanism and clinical stage), laboratory (viral load (VL), baseline CD4 and existence of prior mutations test) and TAR chosen were recorded. VL and CD4, adherence, monitoring, medication-related problems, changes and reasons for changing ART were registered at 24 and 48 weeks. Results: Of 253 patients selected, analytical data was available for 244 and 226 at 24 and 48 weeks, respectively. After 24 weeks, VL was <50 copies/ml in 142 (58.23%) patients and <200 copies/ml in 204 (83.6%). After 48 weeks, the same values were 198 (88.4%) and 217 (96.0%), respectively. Baseline VL above 100,000 copies/ml and poor adhesion to treatment were associated with an increased risk of VF. Thirty per cent of patients switched ART during the first year, mainly because of side effects (31.6%) and simplification of treatment (23.7%). Conclusions: VL became undetectable or under 200 copies/ml in a high percentage of patients starting ART within the first year of treatment. Despite the increasingeffectiveness and tolerability of available drugs, side effects remain as the major reason for changing ART.

Should We Be Testing for Baseline Integrase Resistance in Patients Newly Diagnosed With Human Immunodeficiency Virus?

Background: Current guidelines recommend genotype resistance testing at diagnosis to guide initial selection of antiretroviral therapy (ART). Many standard resistance genotypes exclude testing for resistance to integrase inhibitors ("IR testing"), although this class of drugs is a component of most recommended first-line regimens. Methods: We compared the 96-week clinical outcomes and cost-effectiveness of 2 strategies: no IR testing vs IR testing performed at human immunodeficiency virus (HIV) diagnosis. The base case prevalence of transmitted integrase strand transfer inhibitor (INSTI)-resistant (INSTI-R) virus is estimated at 0.1%. With no IR testing, all patients start dolutegravir (DTG)-based ART after genotype; 12-week suppression rates are 90% (INSTI-susceptible [INSTI-S] virus) and 35% (INSTI-R virus). Those not suppressed at 12 weeks undergo IR testing; if diagnosed with INSTI-R virus, they change to ritonavir-boosted darunavir (DRV/r)-based ART. With IR testing, all patients are diagnosed with INSTI-S/INSTI-R virus prior to ART initiation and start DTG- or DRV/r-based regimens, respectively. Costs include IR tests (175 US dollars [USD]) and ART (41100-44900 USD/year). We examined the impact of key parameters in sensitivity analyses. Results: IR testing resulted in worse clinical outcomes compared to no IR testing and increased costs by 200 USD/person/year. Prevalence of transmitted INSTI-R virus did not affect the favored strategy. No IR testing remained clinically preferred unless DTG suppression of INSTI-R virus was <20% or 96-week DRV/r suppression was >92%. If quality of life was worse with DRV/r- than DTG-based ART, no IR testing was clinically preferred over an even broader range of parameters. Conclusions: In patients with newly diagnosed HIV, IR testing is projected to result in worse outcomes and is not cost-effective. Pretreatment assessment for INSTI resistance should not be recommended in treatment guidelines.

Factors influencing the virological testing of cornea donors.

To assess the influence of donor, environment, and logistical factors on the results of virological testing of blood samples from cornea donors.Data from 670 consecutive cornea donors were analyzed retrospectively. Logistic regression analysis was used to assess the influence of different factors on the results of virological testing of blood samples from cornea donors.The mean annual rate of donors with serology-reactive or not evaluable result was 14.8% (99 of 670) (range 11.9%-16.9%). The cause of donor death by cancer increased the risk of serology-reactive or not evaluable result (P = .0300). Prolonged time between death and post mortem blood removal was associated with a higher rate of serology-reactive or not evaluable result (P < .0001). Mean monthly temperature including warmer months, differentiating between septic and aseptic donors, sex, and donor age had no significant impact on the results of virological testing of blood samples from cornea donors.The cause of donor death by cancer and a prolonged time between death and post mortem blood removal seem to be mainly responsible for serology-reactive or not evaluable result of blood samples from cornea donors. The percentage of discarded corneas caused by serology-reactive or not evaluable result may be reduced by shortening the period of time between death and post mortem blood removal.

Longitudinal follow-up of Zika virus RNA in semen of a traveller returning from Barbados to the Netherlands with Zika virus disease, March 2016

We report the longitudinal follow-up of Zika virus (ZIKV) RNA in semen of a traveller who developed ZIKV disease after return to the Netherlands from Barbados, March 2016. Persistence of ZIKV RNA in blood, urine, saliva and semen was followed until the loads reached undetectable levels. RNA levels were higher in semen than in other sample types and declined to undetectable level at day 62 post onset of symptoms...(AU)

Acute viral hepatitis - should the current screening strategy be modified?

BACKGROUND: The epidemiology of viral hepatitis has changed. Since the introduction of safe and effective vaccines for hepatitis A and B in 1980s, the incidence of acute infections caused by these viruses has been declining in the UK. At the same time, hepatitis E virus (HEV) has been recognised as an increasingly important cause of acute hepatitis, but testing is not widely available. OBJECTIVES: The aim of this study was to establish the viral causes of acute hepatitis, and use that data to modify the current diagnostic algorithm. STUDY DESIGN: A Cognos search was performed to collate subjects tested for HAV, HBV, HCV, HEV, EBV and CMV between June 2010 and December 2012. Information included virological result and their ALT level if done within 5 days from virological testing. RESULTS: From 3462 subjects with suspected acute viral hepatitis, only 25% had biochemical evidence of acute hepatitis (n=854; ALT>100IU/l). The frequency of detection of acute HEV infection (25/409) was over 31-times higher than that of HAV (6/3462), and 7-times higher than that of HBV (24/3462). Most cases of acute HAV, HEV, EBV and CMV infections presented with abnormal ALT levels. Most EBV infections were associated with lymphadenopathy (23/34); in comparison most of CMV infections were not associated with lymphadenopathy (18/22). CONCLUSIONS: HEV screening should be included in the initial testing panel for acute hepatitis and screening at least for HAV and HEV might be limited to those with abnormal ALT levels.

Survey of diagnostic services for genital herpes in fourteen countries in Eastern Europe.

This paper reports survey-based data on the diagnosis and management of genital herpes simplex virus (HSV) infection in 14 countries of the Eastern European Network for Sexual and Reproductive Health (EE SRH). Only 43% of the countries could provide the number of genital HSV cases recorded at national level. Eighty-six percent of countries employed syndromic management in cases of genital ulcer disease. Most countries performed type-specific and/or non-type-specific enzyme immunoassays to detect HSV antibodies. Non-type-specific serology for diagnostic purposes should be actively discouraged. Direct detection methods for HSV, such as PCR, antigen detection and culture, are available in the region, but their usage was extremely low. Their use in Eastern European countries should be actively promoted. The availability of laboratory services must be improved, and countries in the region should implement consensus recommendations for the laboratory diagnosis of genital HSV infections in order to improve clinical practice.

Categorical data analysis in experimental biology.

The categorical data set is an important data class in experimental biology and contains data separable into several mutually exclusive categories. Unlike measurement of a continuous variable, categorical data cannot be analyzed with methods such as the Student's t-test. Thus, these data require a different method of analysis to aid in interpretation. In this article, we will review issues related to categorical data, such as how to plot them in a graph, how to integrate results from different experiments, how to calculate the error bar/region, and how to perform significance tests. In addition, we illustrate analysis of categorical data using experimental results from developmental biology and virology studies.

Completion of national laboratory inventories for wild poliovirus containment --- region of the Americas, March 2010.

In May 1988, the World Health Assembly resolved to eradicate wild poliovirus (WPV) transmission globally. By 2006, transmission of indigenous WPV was eliminated in all but four countries (Afghanistan, India, Nigeria, and Pakistan). In May 1999, the World Health Assembly urged member states to begin the process leading to laboratory containment of WPV. Containment of infectious and potentially infectious WPV materials after eradication is essential to minimize the risk for reintroducing WPV into poliomyelitis-free communities. The staged containment approach begins with a national survey of all biomedical facilities, which alerts facilities to the need for containment, encourages reduction of WPV materials, and develops a national inventory of facilities holding such materials (Phase I). In May 2008, the World Health Assembly reiterated the need for progress in containment and urged polio-free states to complete Phase I. This report describes completion of Phase I by the countries and territories in the World Health Organization (WHO) Region of the Americas during 2001-2010. Of 67,362 biomedical facilities, all 15,541 (23.1%) that were classified as high-risk or medium-risk facilities were surveyed. Of the remaining 51,821 (76.9%) facilities, all classified as low-risk, 44,077 (85.1%) were surveyed; sampling ranged from 12.8% to 100% among countries. After voluntary destruction of some materials during Phase I, a total of 215 facilities in nine countries of the Region of the Americas reported retaining WPV materials as of March 2010. The survey provides a facility registry for use in subsequent steps that will lead to global poliovirus containment.

National laboratory inventories for wild poliovirus containment--Western Pacific region, 2008.

In the future, when wild poliovirus (WPV) transmission is interrupted worldwide, facilities holding WPV materials will represent the only remaining repository of the virus. Maintaining the number of such facilities at a minimum and at an appropriate biosafety standard (laboratory containment) reduces the risk for a facility-associated reintroduction of WPV. In May 1999, the World Health Assembly (WHA) urged all member states to begin the process leading to laboratory containment of WPV. The World Health Organization (WHO) global action plan for laboratory containment of WPV issued in 1999 indicated a staged approach that begins with a national survey of all biomedical facilities (Phase I); the purpose of the survey is to alert institutions and facilities to the need for containment, encourage reduction of WPV materials, and develop a national inventory of facilities holding such materials. The survey and inventory provide a facility database for use in all subsequent steps toward global poliovirus containment. In May 2008, WHA urged all WHO member states to complete Phase I activities outlined in the WHO Global Action Plan for Laboratory Containment of Wild Polioviruses. In the WHO Western Pacific Region (WPR), Phase I surveys of 77,260 laboratories in the 37 countries and areas of WPR were conducted during 1999--2008. A total of 45 laboratories were identified as holding WPV materials in 2008. This report describes completion of Phase I containment activities by WPR countries, and updates a previous report on Phase I completion in the European Region and global progress.

Diagnostic virology practices for respiratory syncytial virus and influenza virus among children in the hospital setting: a national survey.

A survey was sent to the emergency room and laboratory directors of 400 randomly selected US hospitals to assess the diagnostic testing practices for respiratory syncytial virus and influenza virus in children. The results demonstrate that the majority of hospitals routinely perform viral testing for both viruses and use virology testing practices appropriate for the reasons reported for testing.

HPV detection methods.

Given the causal relation between a persistent high-risk human papillomavirus (hrHPV) infection and the development of high-grade cervical intraepithelial neoplasia (CIN) and cervical cancer, hrHPV testing has been advocated in addition to cytology for the detection of clinically relevant cervical lesions. HrHPV testing is thought to improve cervical screening algorithms, the management of women with cytologically equivocal smears, and the management of women treated for high grade CIN. In this chapter we discuss different methods for HPV detection and genotyping and their respective applications.

International standard reagents for HPV detection.

Human papillomavirus is the commonest genital viral infection in healthy sexually active subjects, and the presence of chronic or persistent HPV types in genital cells may constitute a prognostic marker of underlying, or predict future HPV-associated diseases. A variety of novel tests for detecting the presence of oncogenic HPV types in biological specimens have been reported. These are based on the various stages of infection and viral life cycle. HPV infects squamous epithelium with expression of various gene products intimately linked to epithelial cell differentiation. Hence, there are basically three classes of detectable markers directly derived from HPVs: molecular markers based on detection of nucleic acid sequences, serological markers based on detection of antibodies against viral proteins, and cellular markers based on detection of proteins expressed intracellularly, upon either infection or carcinogenesis. The nature of various assays and the development of international standard reagents for qualitative and quantitative assessment of assay performance are outlined. There is an increasing demand to develop standard tools to assess the quality of HPV detection systems, for regulatory and clinical management purposes. International standard reagents for HPV will help defining the analytical sensitivity and specificity of various detection methods, and will allow assuring that laboratory services used to evaluate disease burden, HPV vaccines, and cancer prevention strategies are accurate and comparable worldwide. The advancement of prophylactic vaccine candidates against HPV infections and related diseases stresses the increasing importance of HPV assays in monitoring the impact of HPV vaccination on disease burden.

The analysis and regulation for the dynamics of a temperate bacteriophage model.

The purpose of this paper is to study the asymptotical behavior of a temperate bacteriophage model in chemostat, which was first proposed by Levin et al. [B.R. Levin, F.M. Stewart, L. Chao, Resource-limited growth, competition and predation: A model and experiment studies with bacteria and bacteriophage, Am. Nat. 125 (1977) 3]. Firstly, a classification for the equilibria of the model and their stability are obtained; secondly, sufficient conditions for uniform persistence are obtained; thirdly, sufficient conditions for the global asymptotic behavior are given, and simulations for the model are presented. The theoretical results show that there are more than eight cases for the classification of the model, and that the decrease (increase) of the nutrient concentration or average lytic time (flow rate) is beneficial to the survival of the sensitive cells. Both the simulated and theoretical results show that there is a possibility of switch phenomena or a periodical outburst of the phages and the lysogens, which is caused by the internal factors rather than by some external environment. Finally, the simulation and regulation of the dynamics of the model with experimental data are presented.

Fast, ultrasensitive virus detection using a Young interferometer sensor.

We report the application of an integrated optical Young interferometer sensor for ultrasensitive, real-time, direct detection of viruses. We have validated the sensor by detecting herpes simplex virus type 1 (HSV-1), but the principle is generally applicable. Detection of HSV-1 virus particles was performed by applying the virus sample onto a sensor surface coated with a specific antibody against HSV-1. The performance of the sensor was tested by monitoring virus samples at clinically relevant concentrations. We show that the Young interferometer sensor can specifically and sensitively detect HSV-1 at very low concentrations (850 particles/mL). We have further demonstrated that the sensor can specifically detect HSV-1 suspended in serum. Extrapolation of the results indicates that the sensitivity of the sensor approaches the detection of a single virus particle binding, yielding a sensor of unprecedented sensitivity with wide applications for viral diagnostics.

National laboratory inventory for global poliovirus containment--European Region, June 2006.

In May 1999, the World Health Assembly reaffirmed the commitment of the World Health Organization (WHO) to eradicate poliomyelitis and urged all member states to begin the process leading to the laboratory containment of wild poliovirus (WPV). The WHO global action plan for laboratory containment of WPV begins with a survey of all biomedical facilities (Phase I). The purpose of the survey is to alert institutions and facilities to the need for containment, encourage reduction of WPV materials, and develop a national inventory of facilities holding such materials. The objective of Phase I is to provide a facility database for use in all subsequent steps toward global poliovirus containment. This report describes completion of Phase I containment by the European Region, the first of the six WHO regions to accomplish this goal.

Comparative performance of the Amplicor HIV-1 Monitor Assay versus NucliSens EasyQ in HIV subtype C-infected patients.

In facing global programs for treating HIV-infected patients in the developing countries, there is a real need for viral load assays that are accurate for the local subtypes. The present study was designed to evaluate viral load measurements using the newer version of the NASBA assay in subtype C-infected patients. The performances of this new version, a real-time nucleic acid sequence-based amplification HIV-1 assay (NucliSens EasyQ), were compared to Amplicor HIV-1 Monitor Assay version 1.5 in 79 samples of subtype C-infected patients originating from Ethiopia. Twenty HIV-1 subtype B-infected patients served as a control group. Blood samples from patients in both groups were tested by both assays. The results were compared by a paired, two-tailed Student's t-test. The disparity between the results of the two viral load assays was highly significant in subtype C samples (P = 0.005), such that in the vast majority, higher values of viral load were obtained by the Amplicor assay. However, no differences between the two assays were found in subtype B samples (P = 0.77). CD4 measurements were available for 78 samples of subtype C-infected patients. Of these, a CD4-to-viral load discrepancy (CD4

Quantitation of cytomegalovirus DNA by real-time polymerase chain reaction in peripheral blood specimens of patients with solid organ transplants: comparison with end-point PCR and pp65 antigen test.

The polymerase chain reaction (PCR) for cytomegalovirus (CMV) DNA quantitation provides sensitive and specific data for detecting CMV as well as monitoring the infection and determining the appropriate antiviral strategy. A recently introduced real-time PCR assay for CMV DNA quantitation was applied on 158 peripheral blood leukocytes (PBLs) from 32 liver-transplanted patients with CMV asymptomatic infection and correlated with a commercial quantitative end-point PCR (COBAS AMPLICOR CMV Monitor) and CMV pp65 antigenemia. A good correlation was found between real-time PCR and pp65 antigen test (r2 = 0.691) and the two PCR assays (r2 = 0.761). Real-time PCR data were higher in pre-emptive treated patients (>20 pp65 + positive cells, median CMV DNA value: 3.8 log(10) copies/500,000 PBLs) than in not-treated ones (2.9 logs). According to pp65 levels of 0, 1-10, 11-20, 21-50, 51-100, and >100 positive cells/200,000 PBLs, median CMV DNA by real-time PCR was 2.6, 3.0, 3.6, 4.0, 4.2, and 4.8 logs, respectively, (CMV DNA levels by COBAS AMPLICOR: 2.8, 2.9, 3.8, 3.7, 3.9, and 4.0 logs). For samples with >20 pp65 + cells, real-time PCR gave significantly higher values than in groups with <20 pp65 + cells, whereas the COBAS AMPLICOR results showed a slower progression rate. Dilutions of CMV AD169 strain were used to probe real-time PCR reproducibility (between and intra-assay variability <2%) and sensitivity (100% detection rate at 10 copies/reaction, 28.5% with end-point PCR). In conclusion, real-time PCR significantly improves the study of CMV DNA dynamics due to a more reliable quantitation of CMV DNA for moderate and high DNA level compared to end-point PCR with better sensitivity and specificity. Real-time PCR provides more precise information for evaluating infection progress and assessing antiviral response, simplifying and accelerating the process of producing a reliable quantitation of CMV DNA for clinical purposes.