JOURNAL OF CLINICAL VIROLOGY

Nucleic acid amplification testing using dried blood spots to confirm the diagnosis of HIV-1 in adults
Pinsky BA, Sahoo MK, Manasa J, Makadzange T, Wallis CL, Marins EG, Kumarasamy N, Bartlett JA, Bosch RJ, Israelski D, Katzenstein DA and
The WHO HIV testing algorithm for high prevalence populations recommends the use of three different serologic assays, though this approach may lead to diagnostic misclassification. The study objective was to compare dried blood spot (DBS)-based HIV-1 nucleic acid detection methods to determine their suitability to confirm the diagnosis of HIV-1 in adults generally with suppressed or low-level plasma HIV-1 RNA.
Coinfections and iterative detection of respiratory viruses among 17,689 patients between March 2021 and December 2022 in Southern France
Mantelli C, Colson P, Lesage L, Stoupan D, Chaudet H, Morand A, La Scola B and Boschi C
We aimed to describe coinfections and iterative infections with respiratory viruses diagnosed over a 22-month period in 2021-2022 in public university hospitals of the second largest French city.
Metagenomic characterization of viruses in the serum of children with newly diagnosed cancer
Leijonhufvud G, Soratto TAT, Matos GM, Bajalan A, Eichler-Jonsson C, Gustafsson B, Bogdanovic G, Allander T, Ljungman G and Andersson B
A large cohort of pediatric patients with various forms of childhood cancer was investigated for the presence of viruses using metagenomics. A total of 476 patient samples, collected between 1989 and 2018, were analyzed, representing various pediatric oncological diagnoses and a control group of non-malignant diagnoses.
Parvovirus B19 remains an underestimated pathogen among infections during gestation in Argentina: Insights through the study of symptomatic and asymptomatic pregnant patients and newborns from Córdoba
Colazo Salbetti MB, Boggio G, Dicuatro N, Gudiño AP, Olivera N, Pedranti M, Isa MB, Bertoldi A, Miranda MJ, Rodriguez Lombardi G, Sicilia P, Castro G, Moreno L and Adamo MP
Parvovirus B19 (B19 V) infection during pregnancy can cause adverse fetal outcomes. Our aim was to characterize both clinical and asymptomatic maternal and neonatal cases by studying virological and serological markers of B19 V infection, and to sequence the complete genome of the circulating virus in Argentina.
Opt-out HIV screening in adults attending the Emergency Department of a London teaching hospital
Rachman R, Haque T, Barber TJ, Burns F, Pinto J, Hunter A, Durkin R and Hart J
Opt-out Emergency Department blood borne virus (EDBBV) screening was introduced at the Royal Free Hospital under the NHSEI (NHS England and NHS Improvement) programme to expand opt-out testing in local authority areas with high HIV prevalence. This initiative was part of the "Toward Zero" policy towards ending HIV transmission in England by 2030.
Clinical accuracy of OncoPredict HPV Quantitative Typing (QT) assay on self-samples
Latsuzbaia A, Martinelli M, Giubbi C, Cuschieri K, Elasifer H, Iacobone AD, Bottari F, Piana AF, Pietri R, Tisi G, Odicino F, Cocuzza CE, Arbyn M and
The VALHUDES initiative was established to assess the clinical accuracy of HPV assays to detect cervical precancers using urine and vaginal self-samples compared to cervical clinician-collected samples. Here, the clinical performance of OncoPredict HPV Quantitative Typing (QT) assay (OncoPredict QT) was evaluated.
Detection of yellow fever virus genome in urine following natural infection or vaccination: review of current knowledge 1985-2023
Igloi Z, Pezzi L, Charrel RN and Koopmans M
Yellow fever virus (YFV) is endemic in the (sub)tropical regions of Africa and South America and is prone to cause epidemics. Molecular testing of YFV by reverse transcription-polymerase chain reaction (RT-PCR) was recently adopted by WHO using blood. Urine is a non-invasive diagnostic specimen which has been proven to be useful in diagnosing several flavivirus infections. Until now, systematic data on the usefulness of urine in YFV molecular diagnostics was lacking.
International external quality assessment study for detection of monkeypox virus by PCR supporting laboratory preparedness during the 2022-2023 mpox outbreak and beyond
Ehmann R, Donoso Mantke O, McCulloch E, Yousef A, Ricketts A, Staines H, Bugert JJ, Wölfel R and Niesters HGM
Diagnostic capabilities and correspondent External Quality Assessments (EQA) are key for outbreak preparedness. To support diagnostic facilities with a quality assessment of newly established monkeypox virus (MPXV) molecular diagnostic workflows, Quality Control for Molecular Diagnostics (QCMD) and the Bundeswehr Institute of Microbiology (IMB) piloted an international EQA study conducting four challenges from autumn 2022 to summer 2023 during the global mpox outbreak.
Performance evaluation of the Abbott Alinity Hepatitis C antigen next assay in a US urban emergency department population
Prostko J, Rothman R, Hsieh YH, Pearce S, Kilbane M, McAuley K, Frias E, Taylor R, Ali H, Buenning C, Grieshaber J, Bedrava J and Daghfal D
HCV antibody assays have been used to screen for HCV, but confirmation of acute infection is dependent on RNA or core antigen testing. The aim of the study was to compare the performance of five HCV test methods, including RNA testing, on a US emergency department population.
Standardization and evaluation of an in-house ELISA for the detection of rabies antibody in a tertiary care centre in South India
Thangavelu K, Daniel HD, Kannangai R, Abraham AM, Velladurai SK and Mammen DS
Rapid fluorescent focus inhibition test (RFFIT), a neutralization-based assay for detecting rabies antibodies, is the gold standard. The National Action Plan for Dog Mediated Rabies Elimination (NAPRE) is a national program that strategizes the establishment of enzyme-linked immunosorbent assays (ELISA) to detect rabies antibodies.
Validation, implementation and quality control of a Torque Teno Virus qPCR in a multinational clinical trial
Gore EJ, Gard L, Bourgeois P, Kulifaj D, McCulloch E, Spezia PG, Niesters HGM, Maggi F, Bond G, Van Leer-Buter C and
Immunosuppressive medication after organ transplantation is usually dosed through therapeutic drug monitoring. Trough levels of antirejection medication however, do not adequately predict rejection or infections. The TTVguideIT trial is a multinational clinical trial evaluating the safety of Torque Teno Virus (TTV) load assessed by qPCR, as an alternative to trough level tacrolimus dosing.
High-risk HPV mRNA testing on self-samples offered to those who do not attend for organised cervical screening - real world data from the Dumfries and Galloway region in Scotland
Forson W, Bhatia R, Currie H, Elasifer H, Connor L, Wilson A and Cuschieri K
HPV self sampling can act as a tool to engage women in cervical screening and population based studies can inform optimal implementation of this approach.
Genomic evolution of influenza during the 2023-2024 season, the johns hopkins health system
Yunker M, Villafuerte DA, Fall A, Norton JM, Abdullah O, Rothman RE, Fenstermacher KZJ, Morris CP, Pekosz A, Klein E and Mostafa HH
Influenza, a human disease caused by viruses in the Orthomyxoviridae family, is estimated to infect 5% -10 % of adults and 20% -30 % of children annually. Influenza A (IAV) and Influenza B (IBV) viruses accumulate amino acid substitutions (AAS) in the hemagglutinin (HA) and neuraminidase (NA) proteins seasonally. These changes, as well as the dominating viral subtypes, vary depending on geographical location, which may impact disease prevalence and the severity of the season. Genomic surveillance is crucial for capturing circulation patterns and characterizing AAS that may affect disease outcomes, vaccine efficacy, or antiviral drug activities. In this study, whole-genome sequencing of IAV and IBV was attempted on positive remnant clinical samples (587) collected from 580 patients between June 2023 and February 2024 in the Johns Hopkins Health System (JHHS). Full-length HA segments were obtained from 424 (72.2 %) samples. H1N1pdm09 (71.7 %) was the predominant IAV subtype, followed by H3N2 (16.7 %) and IBV-Victoria clade V1A.3a.2 (11.6 %). Within H1N1pdm09 HA sequences, the 6B1A.5a.2a.1 (60.5 %) clade was the most represented. Full-length NA segments were obtained from 421 (71.7 %) samples. Within H1N1pdm09 and IBV, AAS previously proposed to change susceptibility to NA inhibitors were infrequently detected. Phylogeny of HA and NA demonstrated heterogeneous HA and NA H1N1pdm09 and IBV subclades. No significant differences were observed in admission rates or use of supplemental oxygen between different subtypes or clades. Influenza virus genomic surveillance is essential for understanding the seasonal evolution of influenza viruses and their association with disease prevalence and outcomes.
Hepatitis B virus (HBV) viremia despite tenofovir disoproxil fumarate-containing antiretroviral therapy in persons with HBV/HIV coinfection
Ryan P, Odegard E, Meeds H, Lartey M, Ganu VJ, Tachi K, Yang H, Ojewale O, Boamah I, Obo-Akwa A, Antwi K, Anderson PL, Blackard JT and Kwara A
The goal of treatment of hepatitis B virus (HBV) and human immunodeficiency virus (HIV) coinfection is suppression of both viruses; yet incomplete HBV suppression on tenofovir (TFV) disoproxil fumarate (TDF)-based antiretroviral therapy (ART) is common. This study investigated TFV resistance-associated mutations (RAMs) in individuals with HBV/HIV coinfection with viremia on TDF/lamivudine (3TC)-containing ART.
What the pox? Review of poxviruses affecting humans
Hata DJ, Powell EA, Starolis MW and Realegeno SE
The re-emergence of human mpox with the multi-country outbreak and a recent report of borealpox (previously Alaskapox) resulting in one death has heightened awareness of the significance of the Poxviridae family and their zoonotic potential. This review examines various poxviruses affecting humans, with discussion of less commonly encountered Poxviridae members, including pathogenesis, epidemiology, and diagnostic methods. Poxvirus treatment is beyond the intended scope of this review and will not be discussed.
Influenza C virus in U.S. children with acute respiratory infection 2016-2019
Sederdahl BK, Weinberg GA, Campbell AP, Selvarangan R, Schuster JE, Lively JY, Olson SM, Boom JA, Piedra PA, Halasa NB, Stewart L, Szilagyi PG, Balasubramani GK, Sax T, Martin JM, Hickey RW, Michaels MG, Williams JV and
Influenza C virus (ICV) is an orthomyxovirus related to influenza A and B, yet due to few commercial assays, epidemiologic studies may underestimate incidence of ICV infection and disease. We describe the epidemiology and characteristics of ICV within the New Vaccine Surveillance Network (NVSN), a Centers for Disease Control and Prevention (CDC)-led network that conducts population-based surveillance for pediatric acute respiratory illness (ARI). Nasal or/combined throat swabs were collected from emergency department (ED) or inpatient ARI cases, or healthy controls, between 12/05/2016-10/31/2019 and tested by molecular assays for ICV and other respiratory viruses. Parent surveys and chart review were used to analyze demographic and clinical characteristics of ICV+ children. Among 19,321 children tested for ICV, 115/17,668 (0.7 %) ARI cases and 8/1653 (0.5 %) healthy controls tested ICV+. Median age of ICV+ patients was 18 months and 88 (71.5 %) were ≤36 months. Among ICV+ ARI patients, 40 % (46/115) were enrolled in the ED, 60 % (69/115) were inpatients, with 15 admitted to intensive care. Most ICV+ ARI patients had fever (67.8 %), cough (94.8 %), or wheezing (60.9 %). Most (60.9 %) ARI cases had ≥1 co-detected viruses including rhinovirus, RSV, and adenovirus. In summary, ICV detection was rarely associated with ARI in children, and most ICV+ patients were ≤3 years old with co-detected respiratory viruses.
Opportunities and challenges for the U.S. laboratory response to highly pathogenic avian influenza A(H5N1)
Pinsky BA and Bradley BT
On March 25, 2024 an outbreak of highly pathogenic avian influenza (HPAI) A H5N1 was identified in dairy cows across multiple farms in the United States. Zoonotic cases originating in individuals with close contact to infected herds and poultry flocks have been subsequently identified. Spillover events such as this raise the specter of recent pandemics including COVID-19 and Mpox and may lead clinical laboratories to assess their capacity for diagnosis of HPAI H5N1. In this review, we detail the origins of the H5N1 clade 2.3.4.4b outbreak as well as the existing capacity to identify HPAI H5N1 as influenza A virus by commercially available assays. Furthermore, we highlight the absence of commercially available influenza A H5 subtyping assays and limitations associated with the current 510(k)-cleared assay. This outbreak also serves as an early opportunity to assess the new and unknown regulatory challenges faced by laboratory-developed tests in light of the FDA's final rule on in vitro diagnostic devices. National agencies along with public health and clinical laboratories all serve an essential role in the response to HPAI H5N1. To most effectively utilize each group's strength requires open communication and willingness to embrace novel approaches.
Performance evaluation of the quantitative assay Adenovirus ELITe MGB® Kit on EDTA-plasma, using the ELITe BeGenius® system
Fries E, Kalmeijer T, Spaans L, Rakké M, Schuurman R and De Groot-Mijnes JDF
Adenovirus infections constitute an important cause of morbidity and mortality after hematopoietic stem cell transplantation. Detection and monitoring of adenovirus in EDTA-plasma by real-time quantitative PCR is a sensitive tool for identification and management of patients at risk of a potentially fatal infection.
Circulating serotypes and genotypes of dengue virus during the 2023 outbreak in Eastern Nepal
Bharati N, Dumre SP, Shah Y, Nabesima T, Dhimal M, Pandey S, Kapandji M, Takamatsu Y, Urano T, Pandey BD, Morita K, Ngwe Tun MM and Pandey K
Dengue virus (DENV) is one of the most significant mosquito-borne diseases in Nepal. In 2023, DENV outbreaks began in Eastern Nepal, near the border with India, and rapidly spread nationwide. The study aims to describe the outbreak's epidemiological pattern, laboratory characteristics, DENV serotypes, and genotypes. A hospital-based cross-sectional study was conducted in four hospitals in Jhapa, Eastern Nepal, in 2023. Acute serum samples were obtained from dengue suspected patients within 7 days of illness and subjected to virus isolation, conventional and real-time polymerase chain reaction (RT-PCR), and phylogenetic analysis. Out of 60 samples, 42 (70 %), 11 (18.3 %) and 7 (11.7 %) were primary, secondary and non-dengue infection, respectively. Among 53 dengue confirmed patients, 46 (86.7 %) were positive for NS1 and 12 (22.6 %) were positive for both NS1 and IgM. Out of 42 dengue isolates, a new clade of the cosmopolitan genotype of DENV-2 was the most prevalent (28, 66.7 %), followed by genotype III of DENV-3 (11, 26.2 %) and genotype V of DENV-1 (3, 7.1 %). Genotype III of DENV-3 was first introduced in 2022-2023 in Nepal. Phylogenetic analysis of the E gene revealed the DENV-2 isolates from Nepal had 98 % homologous nucleotide similarity with the strains from India and Bangladesh. To our knowledge, this is the first report of circulating serotypes and genotypes of DENV in Jhapa. Integrating molecular findings into the dengue control plan can enhance surveillance efforts, monitor disease trends, and implement proactive measures to reduce the burden of dengue and prevent fatalities in future outbreaks.
What have we learned from animal studies of immune responses to respiratory syncytial virus infection?
Drysdale SB, Thwaites RS, Price J, Thakur D, McGinley J, McPherson C, Öner D, Aerssens J, Openshaw PJ, Pollard AJ and
Respiratory syncytial virus (RSV) is a common cause of severe respiratory tract infection at the extremes of age and in vulnerable populations. However, it is difficult to predict the clinical course and most infants who develop severe disease have no pre-existing risk factors. With the recent licencing of RSV vaccines and monoclonal antibodies, it is important to identify high-risk individuals in order to prioritise those who will most benefit from prophylaxis. The immune response to RSV and the mechanisms by which the virus prevents the establishment of immunological memory have been extensively investigated but remain incompletely characterised. In animal models, beneficial and harmful immune responses have both been demonstrated. While only chimpanzees are fully permissive for human RSV replication, most research has been conducted in rodents, or in calves infected with bovine RSV. Based on these studies, components of innate and adaptive immune systems, cytokines, chemokines and metabolites, and specific genetic and transcriptomic signatures are identified as potential predictive indicators of RSV disease severity. These findings may inform the development of future human studies and contribute to the early identification of patients at high risk of severe infection. This narrative review summarises the factors involved in the immune response to RSV infection in these models and highlights the relationship between potential biomarkers and disease severity.
Investigation of suspected false positive norovirus results on a syndromic gastrointestinal multiplex molecular panel
Caza M, Kuchinski K, Locher K, Gubbay J, Harms M, Goldfarb DM, Floyd R, Kenmuir E, Kalhor M, Charles M, Prystajecky N and Wilmer A
Suspected false positive norovirus results occurred after introduction of the BioFire® FilmArray® Gastrointestinal panel (BF-GIP) into 6 laboratory sites, in British Columbia, Canada.