Mark M. Manak

Prospective Study of Acute HIV-1 Infection in Adults in East Africa and Thailand

BACKGROUNDnAcute human immunodeficiency virus type 1 (HIV-1) infection is a major contributor to transmission of HIV-1. An understanding of acute HIV-1 infection may be important in the development of treatment strategies to eradicate HIV-1 or achieve a functional cure.nnnMETHODSnWe performed twice-weekly qualitative plasma HIV-1 RNA nucleic acid testing in 2276 volunteers who were at high risk for HIV-1 infection. For participants in whom acute HIV-1 infection was detected, clinical observations, quantitative measurements of plasma HIV-1 RNA levels (to assess viremia) and HIV antibodies, and results of immunophenotyping of lymphocytes were obtained twice weekly.nnnRESULTSnFifty of 112 volunteers with acute HIV-1 infection had two or more blood samples collected before HIV-1 antibodies were detected. The median peak viremia (6.7 log10 copies per milliliter) occurred 13 days after the first sample showed reactivity on nucleic acid testing. Reactivity on an enzyme immunoassay occurred at a median of 14 days. The nadir of viremia (4.3 log10 copies per milliliter) occurred at a median of 31 days and was nearly equivalent to the viral-load set point, the steady-state viremia that persists durably after resolution of acute viremia (median plasma HIV-1 RNA level, 4.4 log10 copies per milliliter). The peak viremia and downslope were correlated with the viral-load set point. Clinical manifestations of acute HIV-1 infection were most common just before and at the time of peak viremia. A median of one symptom of acute HIV-1 infection was recorded at a median of two study visits, and a median of one sign of acute HIV-1 infection was recorded at a median of three visits.nnnCONCLUSIONSnThe viral-load set point occurred at a median of 31 days after the first detection of plasma viremia and correlated with peak viremia. Few symptoms and signs were observed during acute HIV-1 infection, and they were most common before peak viremia. (Funded by the Department of Defense and the National Institute of Allergy and Infectious Diseases.).

Laboratory evaluation of rapid test kits to detect hepatitis C antibody for use in predonation screening in emergency settings.

BACKGROUND: Emergency whole blood transfusion is a lifesaving procedure employed on modern battlefields. Rapid device tests (RDTs) are frequently used to mitigate transfusion‐transmitted infection risks.

Transfusion-transmitted human T-lymphotropic virus Type I infection in a United States military emergency whole blood transfusion recipient in Afghanistan, 2010.

The United States introduced human T‐lymphotropic virus Type I (HTLV‐I) screening of blood donors in 1988. The US military uses freshly collected blood products for life‐threatening injuries when available stored blood components in theater have been exhausted or when these components are unsuccessful for resuscitation. These donors are screened after donation by the Department of Defense (DoD) retrospective testing program. All recipients of blood collected in combat are tested according to policy soon after and at 3, 6, and 12 months after transfusion.

Tired of the same old grind in the new genomics and proteomics era

Abstract New discoveries in life sciences depend on accurate analysis of biomolecules, which in turn depends on the extraction of high-quality molecules in high quantities from the tissues of plants, animals or microorganisms. The extraction process for hard-to-lyse cells and tissues has been a bottleneck in the path to discovery for many years. This review describes extraction methods currently in use, and compares them to a newly developed, automated process involving patented pressure cycling technology (PCT). The PCT sample preparation system (SPS) uses an instrument capable of rapid, temperature-controlled pressure cycling between ambient and high pressures, and single-use sample tubes containing a ram and a lysis disk. The quality and quantity of nucleic acid and protein prepared by the PCT SPS method are comparable to the older methods, whereas ease and safety of processing, reproducibility, speed and control are enhanced.

The External Quality Assurance Oversight Laboratory (EQAPOL) proficiency program for IFN-gamma enzyme-linked immunospot (IFN-γ ELISpot) assay.

The interferon-gamma enzyme-linked immunospot (IFN-γ ELISpot) assay has been developed and used as an end-point assay in clinical trials for infectious diseases and cancer to detect the magnitude of antigen-specific immune responses. The ability to compare data generated by different laboratories across organizations is pivotal to understand the relative potency of different therapeutic and vaccine strategies. We developed an external proficiency program for the IFN-γ ELISpot assay that evaluates laboratory performance based on five parameters: timeliness for data reporting; ability to handle cellular samples; detection of background (non-specific) responses; accuracy to consensus of the results; and precision of the measurements. Points are awarded for each criterion, and the sum of the points is used to determine a numeric and adjectival performance rating. Importantly, the evaluation of the accuracy to the consensus mean for the detection of antigen-specific responses using laboratory-specific procedures informs each laboratory and its sponsor on the degree of concordance of its results with those obtained by other laboratories. This study will ultimately provide the scientific community with information on how to organize and implement an external proficiency program to evaluate longitudinally the performance of the participating laboratories and, therefore, fulfill the requirements of the GCLP guidelines for laboratories performing end-point IFN-γ ELISpot assay for clinical trials.

Identification of acute HIV-1 infection by Hologic Aptima HIV-1 RNA Qualitative Assay

ABSTRACT The Hologic Aptima HIV-1 Qualitative RNA assay was used in a rigorous screening approach designed to identify individuals at the earliest stage of HIV-1 infection for enrollment into subsequent studies of cellular and viral events in early infection (RV 217/Early Capture HIV Cohort [ECHO] study). Volunteers at high risk for HIV-1 infection were recruited from study sites in Thailand, Tanzania, Uganda, and Kenya with high HIV-1 prevalence rates among the populations examined. Small-volume blood samples were collected by finger stick at twice-weekly intervals and tested with the Aptima assay. Participants with reactive Aptima test results were contacted immediately for entry into a more comprehensive follow-up schedule with frequent blood draws. Evaluation of the Aptima test prior to use in this study showed a detection sensitivity of 5.5 copies/ml (50%), with all major HIV-1 subtypes detected. A total of 54,306 specimens from 1,112 volunteers were examined during the initial study period (August 2009 to November 2010); 27 individuals were identified as converting from uninfected to infected status. A sporadic reactive Aptima signal was observed in HIV-1-infected individuals under antiretroviral therapy. Occasional false-reactive Aptima results in uninfected individuals, or nonreactive results in HIV-1-infected individuals not on therapy, were observed and used to calculate assay sensitivity and specificity. The sensitivity and specificity of the Aptima assay were 99.03% and 99.23%, respectively; positive and negative predictive values were 92.01% and 99.91%, respectively. Conversion from HIV-1-uninfected to -infected status was rapid, with no evidence of a prolonged period of intermittent low-level viremia.

HODGKIN'S DISEASE AND HUMAN HERPESVIRUS-6: A MODEL FOR STUDIES OF NEW AETIOLOGICAL AGENTS

Although clinical and epidemiological evidence support the role of infectious agents in the aetiology of Hodgkin’s disease (HD), it has not been possible to identify a single pathogen that fits with the epidemiology of this disease. Epstein-Barr virus (EBV) has been linked to HD by a number of serological (Levine et al., 1970; Johannson et al., 1970; Levine et al., 1971; Hesse et al., 1977; Mueller et al., 1989) and virological (Weiss et al., 1989; Bignon et al., 1990; Jarrett et al., 1991; Pallesen et al., 1991; Ambinder et al., 1993) studies with the strongest EBV-association identified in children and older adults (Jarrett et al., 1991; Ambinder et al., 1993), and this virus remains a serious aetiological candidate for a significant proportion of HD cases. Human herpesvirus-6 (HHV-6), first described in 1986 (Salahuddin et al., 1986), has also been linked to HD by serological (Ablashi et al., 1988; Biberfeld et al., 1988; Clark et al., 1990; Iyengar et al., 1991; Levine et al., 1992a) and virological (Torelli et al., 1991; Gompels et al., 1993) studies. This report will summarise the data suggesting an aetiological association, review relevant studies to place these findings in perspective, and discuss possible approaches to the identification of other candidate aetiological agents based on the studies of these two herpesviruses.