Rapid Antigen Detection Assays Research Articles

Rapid detection of human influenza A viruses by HFman probe-based loop-mediated isothermal amplification assays

Since China abandoned the zero-COVID policy at the end of 2022, a wave of severe Flu pandemic emerged in China. Rapid and accurate diagnosis of Influenza A virus (IAV) is critical for clinical management and therapeutic decision-making of patients with fever. Here, we reported a novel IAV HF-LAMP assay, which can be performed with purified RNA or directly using clinical samples. The assays with purified RNA and clinical samples have high sensitivity with limit of detection (LOD) of 9.6 copies/reaction, 9900 copies/mL, and short sample-to-answer times of 36 and 50 min, respectively. Both assays showed high specificity and significantly higher IAV detection rate than the rapid antigen detection (RAD) assays. Furthermore, we found the vast majority (91.2 %) of children with fever during the pandemic were infected by IAV, and current IAV infection has a very narrow detectable window. The novel IVA HF-LAMP assays will provide robust tools to facilitate early diagnosis of IAV infection in current and future seasonal influenza epidemics.

Evaluation of a Rapid Urine Antigen Detection Assay as a Point-of-Care Test in the Diagnosis of Community-Acquired Pneumonia.

Introduction Community-acquired pneumonia (CAP) is among the most common public health problems encountered throughout the world. CAP is a frequent cause of lower respiratory tract infections among children and geriatric-age persons. The etiology of CAP is complex but generally involves infection with bacteria likeStreptococcus pneumoniae (S. pneumoniae), which is the most common cause of CAP. The underdiagnosis of CAP due to the limitations of conventional culture methods could be responsible for severe morbidity and mortality, especially among susceptible populations.We evaluated the usefulness of a rapid immunochromatographic test (BinaxNOW™, Abbott, Chicago, IL) that detects S. pneumoniae through a rapid urine antigen test (RUAT) as a point-of-care (POC) diagnostic method in the early detection of CAP. Methods A prospective study was conducted in a university-affiliated teaching hospital between January 2019 and September 2019 (nine months). The study recruited 300 inpatients who revealed signs and symptoms associated with pneumonia. The study was approved by the institutional ethics committee, and all participants provided their voluntary informed consent.Laboratory evaluationincluded the collection of sputum samples, which were processed for Gram stain and routine culture. Five milliliters of blood were collected from all the subjects for carrying out a blood culture. A urine sample was collected from each participant for the detection of S. pneumoniae through the point-of-care urinary antigen test. Results Of the 300 patients diagnosed with CAP, the S. pneumoniae RUAT was positive in 110 out of 140 cases of pneumococcal pneumoniae (78.57%). The RUAT results were positive for 20 (66.6%) out of 30 bacteremic patients and for 90 (81.8%) out of 110 patients positive for sputum culture. The RUAT was positive in 10 out of 20 cases of pneumonia with an unknown microbial etiology. The overall sensitivity (78.57%), specificity (100%), positive predictive value (100%), negative predictive value (98.88%), and accuracy (90%) of the RUAT were similar to sputum culture results. Conclusion The RUAT has shown comparable efficacy with sputum culture and therefore can be used as a complementary approach to conventional methods in the early diagnosis of CAP caused byS. pneumoniae.Due toits ease of use andrapid results, it could be incorporated as a POC diagnostic test.

Evaluation of a Rapid Immunochromatographic Middle East Respiratory Syndrome Coronavirus Antigen Detection Assay

Abstract Middle East respiratory syndrome coronavirus (MERS-CoV) infection in humans has a high mortality of >30%. Dromedaries are the reservoir of MERS-CoV and the main source of human infections. However, MERS-CoV infections in dromedaries are usually subclinical. Rapid diagnosis of MERS-CoV infection in these animals is important in preventing camel-to-human transmission of the virus. The possible cross-reactivity of a previously reported rapid nucleocapsid protein-based antigen detection assay for MERS-CoV was examined with different CoVs, including Tylonycteris bat CoV HKU4, dromedary camel CoV UAE-HKU23, human CoV-229E, human CoV-OC43, severe acute respiratory syndrome CoV-2 and rabbit CoV HKU14, where none of them showed false-positive results. The assay was further validated using quantitative real-time reverse transcription-polymerase chain reaction–confirmed MERS-CoV–positive and MERS-CoV–negative dromedary nasal samples collected in Dubai, the United Arab Emirates, which showed that the rapid antigen detection assay has a specificity of 100% and sensitivity of 91.7%.

Clinical sensitivity of rapid antigen test during a COVID-19 outbreak in Taipei, May to June 2021

Background/purposeThis population-based study aimed to compare the accuracy of Rapid antigen detection (RAD) and reverse transcription-polymerase chain reaction (RT-PCR) assays for diagnosing individuals infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the COVID-19 outbreak in Taipei, from May to June 2021.MethodsIn response to the outbreak of COVID-19 in mid-May 2021, Taipei City Hospital set up 12 citywide proactive community testing (PCT) stations for early identification of infected individuals from May 17 to June 20, 2021. Individuals with RAD positivity were isolated and later confirmed by RT-PCR. The c-statistic value was estimated to indicate the level of diagnostic accuracy of RAD tests.ResultsOf the 33,798 individuals who were evaluated for SARS-CoV-2 infection, 4.4% tested positive for RAD. There was a moderate concordance (kappa = 0.67) between the RAD tests and RT-PCR assay for identifying infectious individuals. The c-statistic value of the RAD test for the diagnosis of SARS-CoV-2 infection was 0.8. There was a positive linear trend between the accuracy of the RAD tests and the prevalence of SARS-CoV-2 infection in the study population (β = 0.04; p = .03). As the cycle threshold value decreased, the sensitivity rate of the RAD tests increased (p < .001). After implementation of the PCT program, the prevalence of COVID-19 decreased from 8.4% to 3.3% (p < .001).ConclusionProactive community testing for SARS-CoV-2 infection using RAD tests could rapidly identify and quarantine the most infectious patients in the early phase of COVID-19 outbreak.

From more testing to smart testing: data-guided SARS-CoV-2 testing choices, the Netherlands, May to September 2020.

BackgroundSARS-CoV-2 RT-PCR assays are more sensitive than rapid antigen detection assays (RDT) and can detect viral RNA even after an individual is no longer infectious. RDT can reduce the time to test and the results might better correlate with infectiousness.AimWe assessed the ability of five RDT to identify infectious COVID-19 cases and systematically recorded the turnaround time of RT-PCR testing.MethodsSensitivity of RDT was determined using a serially diluted SARS-CoV-2 stock with known viral RNA concentration. The probability of detecting infectious virus at a given viral load was calculated using logistic regression of viral RNA concentration and matched culture results of 78 specimens from randomly selected non-hospitalised cases. The probability of each RDT to detect infectious cases was calculated as the sum of the projected probabilities for viral isolation success for every viral RNA load found at the time of diagnosis in 1,739 confirmed non-hospitalised COVID-19 cases.ResultsThe distribution of quantification cycle values and estimated RNA loads for patients reporting to drive-through testing was skewed to high RNA loads. With the most sensitive RDT (Abbott and SD Biosensor), 97.30% (range: 88.65–99.77) of infectious individuals would be detected. This decreased to 92.73% (range: 60.30–99.77) for Coris BioConcept and GenBody, and 75.53% (range: 17.55–99.77) for RapiGEN. Only 32.9% of RT-PCR results were available on the same day as specimen collection.ConclusionThe most sensitive RDT detected infectious COVID-19 cases with high sensitivity and may considerably improve containment through more rapid isolation and contact tracing.

An optimized stepwise algorithm combining rapid antigen and RT-qPCR for screening of COVID-19 patients.

Background & aimWe investigated the combination of rapid antigen detection (RAD) and RT-qPCR assays in a stepwise procedure to optimize the detection of COVID-19.MethodsFrom August 2020 to November 2020, 43,399 patients were screened in our laboratory for COVID-19 diagnostic by RT-qPCR using nasopharyngeal swab. Overall, 4,691 of the 43,399 were found to be positive, and 200 were retrieved for RAD testing allowing comparison of diagnostic accuracy between RAD and RT-qPCR. Cycle threshold (Ct) and time from symptoms onset (TSO) were included as covariates.ResultsThe overall sensitivity, specificity, PPV, NPV, LR-, and LR+ of RAD compared with RT-qPCR were 72% (95%CI 62%–81%), 99% (95% CI95%–100%), 99% (95%CI 93%–100%), and 78% (95%CI 70%–85%), 0.28 (95%CI 0.21–0.39), and 72 (95%CI 10–208) respectively. Sensitivity was higher for patients with Ct ≤ 25 regardless of TSO: TSO ≤ 4 days 92% (95%CI 75%–99%), TSO > 4 days 100% (95%CI 54%–100%), and asymptomatic 100% (95%CI 78–100%). Overall, combining RAD and RT-qPCR would allow reducing from only 4% the number of RT-qPCR needed.ConclusionsThis study highlights the risk of misdiagnosing COVID-19 in 28% of patients if RAD is used alone. A stepwise analysis that combines RAD and RT-qPCR would be an efficient screening procedure for COVID-19 detection and may facilitate the control of the outbreak.

Development of highly sensitive and rapid antigen detection assay for diagnosis of COVID-19 utilizing optical waveguide immunosensor.

Development of highly sensitive and rapid antigen detection assay for diagnosis of COVID-19 utilizing optical waveguide immunosensor.

Monoclonal antibody against VP0 recognizes a broad range of human parechoviruses

Parechoviruses (PeVs) are common viruses that cause mild gastrointestinal or respiratory symptoms to severe central nervous system infections. In infants, parechovirus infection is one of the leading causes of life-threatening viral disease. High-quality antibodies with broad binding specificities are essential to improve accurate parechovirus diagnosis in diagnostic laboratories. Such antibodies have potential in the development of rapid antigen detection assay against PeVs. In the present study, VP4 and VP2 genes from human parechovirus A1 (PeV-A1) were cloned and VP0 fusion protein produced to develop monoclonal antibodies against PeVs. Two pan-parechovirus antibodies, one IgG and one IgM isotype, were isolated. The properties of IgG1/κ monoclonal (designated as Mab-PAR-1) was studied further. Mab-PAR-1 was shown to be functional in western blot against denatured recombinant protein and viral particles. In immunofluorescence assay, the antibody tested positive for nineteen PeV-A1 isolates while showing no cross-reactivity to fourteen entero- and rhinovirus types. In addition, Mab-PAR-1 showed positive reactivity against five other cultivable parechovirus types 2−6. A unique Mab-PAR-1 epitope located in the junction of the three capsid proteins VP0, VP1, and VP3 was identified using a peptide library screen. This study demonstrates that PeV-A1-VP0 protein is functional antigen for developing monoclonal antibody for diagnosis of broad range of parechovirus infections.

Comparison of five SARS-CoV-2 rapid antigen detection tests in a hospital setting and performance of one antigen assay in routine practice: a useful tool to guide isolation precautions?

BackgroundIn a hospital setting, there is a need for rapid detection of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) to guide isolation measures and targeted admission.AimTo evaluate the diagnostic performance of five SARS-CoV-2 rapid nucleocapsid protein antigen detection (RAD) assays (Biosynex, Biotical, Orient Gene, Panbio and SD Biosensor), and describe the performance and impact of implementation of the SD Biosensor assay in an emergency department.MethodsSensitivity and specificity of the five RAD assays were analysed on 100 respiratory samples: 60 real-time reverse transcriptase polymerase chain reaction (rRT-PCR)-confirmed SARS-CoV-2-positive samples, 24 SARS-CoV-2 RNA-negative samples and 16 samples positive for other respiratory pathogens. The manufacturer's protocol was adapted to validate the antigen tests on transport media used for rRT-PCR in the authors' routine practice. The SD Biosensor RAD assay was implemented as a screening method for rapid diagnosis and targeted admission.FindingsSensitivity of the five RAD assays ranged from 88.9% to 100% for samples with cycle threshold values <26, and specificity ranged from 46.2% to 100%. During the implementation period, 4195 RAD tests were performed. Due to the rapid RAD result, 157 patients were transferred directly to the coronavirus disease 2019 (COVID-19) cohort ward instead of the regular ward (N=47) or the temporary COVID-19 ward (N=110).ConclusionThe SD Biosensor, Biotical and Panbio SARS-CoV-2 antigen tests showed acceptable overall performance, and identified the majority of contagious patients. In the context of high prevalence of SARS-CoV-2, RAD tests can be used as a rapid screening tool to guide infection prevention measures and aid targeted admission.

The Panbio COVID-19 Ag Rapid Test: Which Performances are for COVID-19 Diagnosis?

Rapid antigen detection assays are promising tools for the diagnosis of COVID-19. We assess the performances of the Panbio COVID-19 Ag Rapid Test. The Panbio COVID-19 Ag Rapid test was compared to a reference RT-PCR performed on the same nasopharyngeal swab (NPS). Overall, 81 NPS were tested retrospectively and 330 healthcare workers (HCWs) were tested prospectively. Retrospective analyze. Of the 48 SARS-CoV-2 positive NPS, 19 (39.6%) were found positive with the Panbio COVID-19. There was no cross-reactivity with SARS-CoV-2 negative NPS. The Kappa value was 0.459. Prospective analyze. The prevalence of COVID-19 was 26.1% in symptomatic HCWs. The overall sensitivity and specificity of the Panbio COVID-19 were 47.2% and 100.0% respectively. The sensitivity was 55.2% and 14.3% in those tested within and after 4 days of diseases respectively. The Panbio COVID-19 Ag Rapid test displays low performance for the identification of SARS-CoV-2 infected patients.

A comparative study: Results of rapid antigen detection assay and real time RT-PCR for diagnosis of COVID-19 in a tertiary care hospital

Introduction: December 2019, witnessed the emergence of a novel coronavirus in human population. Later named as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has since spread across the globe and results into coronavirus disease 2019 (COVID-19) pandemic. Objective: The objective of this study was to evaluate COVID-19 infection positivity rate, its age and gender wise prevalence and to compare the results of rapid antigen detection test with gold standard real time RT-PCR for early detection of SARS-CoV-2 virus. Methods: An observational study was conducted in the Department of Microbiology and Molecular Biology of Krishna Institute of Medical Sciences, Secunderabad, Telangana. Results: A total of 208 patients’ specimens (nasopharyngeal swab) were compared within a period of one and half month. Infection positivity rate was observed as 26.92% (n=56). Our study revealed that among all 56 COVID-19 positive patients, 60.71% (n=34) were symptomatic whereas 39.2% (n=22) were asymptomatic. Conclusion: We conclude that after comparing with real time RT-PCR, antigen detection assay is not as sensitive as molecular test especially during the early stages of infection but this has potential to become an important tool for mass screening for COVID-19 and can be considered as fast and economic diagnostic tools in the remote areas where the laboratory facilities for molecular tests are not accessible. Keywords: COVID-19; pandemic; nasopharyngeal swab; rapid antigen detection assay; real time RT-PCR

Cold Weather Viruses.

1. Asif Noor, MD* 2. Theresa Fiorito, MD* 3. Leonard R. Krilov, MD*,† 1. *Department of Pediatrics, Children's Medical Center, NYU Winthrop Hospital, Mineola, NY 2. †Department of Pediatrics, State University of New York, Stony Brook School of Medicine, Stony Brook, NY * Abbreviations: AAP: : American Academy of Pediatrics Adv: : adenovirus CDC: : Centers for Disease Control and Prevention FDA: : Food and Drug Administration hMPV: : human metapneumovirus PCR: : polymerase chain reaction PIV: : parainfluenza virus RSV: : respiratory syncytial virus RV: : rhinovirus Clinicians must learn to identify viral infections in children during the winter months and must practice caution with the use of unnecessary medications in such cases. Recognition of the clinical pattern of viral infection (eg, bronchiolitis) in conjunction with judicious use of viral tests (either office-based immunoassays or newer molecular tests) may assist in epidemiological monitoring, cohorting patients in the hospital, withholding unnecessary therapies, and providing a definitive diagnosis. After completing this article, readers should be able to: 1. Review the epidemiological aspects and clinical signs and symptoms of common cold weather viruses. 2. Recognize situations in which viral testing is indicated. 3. Recognize situations in which treatment is indicated. In early November you are evaluating a 9-month-old boy born at 33 weeks of gestation. The infant presents with 2 days of fevers (101°F–102°F [38.3°C–38.8°C]), copious rhinorrhea, and 1 day of coughing with difficulty breathing. He is otherwise feeding well and has had adequate urination. His 4-year-older sister has an upper respiratory tract infection. On physical examination, the infant has a respiratory rate of 45 breaths/min without chest wall retractions. On auscultation there is good air entry with scattered rhonchi bilaterally. What is the most appropriate next step in management? 1. Obtain respiratory syncytial virus (RSV) and influenza antigen testing. 2. Obtain a chest radiograph to look for focal infiltrate. 3. Provide supportive care with nasal …

Pathology Consultation on Influenza Diagnostics.

Objectives: To describe the strengths and limitations of the available influenza diagnostics, with a focus on rapid antigen detection assays and nucleic acid detection assays. Methods: A case-based presentation is used to illustrate the potential limitations of rapid antigen detection assays for influenza. Results: Influenza is a seasonal illness; estimates attribute influenza to approximately 200,000 hospitalizations and 41,000 deaths in the United States annually. Antigen detection assays for influenza are rapid and convenient, and thus are widely used in a variety of health care settings, even though the sensitivity of these assays may be suboptimal. The United States Food and Drug Administration has recently created new guidelines intended to improve the oversight and performance characteristics of influenza antigen detection assays. Molecular assays, although more costly and complex, are more sensitive and may be designed to simultaneously detect multiple respiratory pathogens within a single assay. Conclusions: Diagnostic assays for influenza can vary greatly with regards to analytical performance characteristics, complexity, turnaround time and cost. This can have important patient care and infection prevention implications.

Quantitative analysis of four rapid antigen assays for detection of pandemic H1N1 2009 compared with seasonal H1N1 and H3N2 influenza A viruses on nasopharyngeal aspirates from patients with influenza

Data on analytical sensitivity of rapid diagnostic assays are important for clinical management of influenza, especially during a pandemic. Four rapid antigen detection assays were compared for detection of pandemic influenza A H1N1 2009, seasonal H1N1 and H3N2 in 96 patients with influenza A infection confirmed by real-time RT-PCR. These rapid antigen tests appeared to have lower sensitivity (55.8%) for the diagnosis of pandemic influenza A H1N1 2009 as compared with seasonal H3N2 (71.0%) or H1N1 (69.4%) influenza infections, a difference that was related to a lower viral load in patients infected with the pandemic influenza A H1N1 2009 virus. The detection limit of these antigen detection tests in clinical specimens was an influenza A M gene copy number of average 1.0×10(7) copies/ml. Significant variations between tests in sensitivity for detection of pandemic influenza A H1N1 2009 (43.4-63.3%) were observed. The Directigen EZ Influenza A+B and the Espline Influenza A+B had comparable sensitivity (63%) and were the most sensitive among the four assays evaluated.

Rapid Detection of Streptococcus pyogenes in Pleural Fluid Samples from Pediatric Patients with Empyema

A total of 120 pleural fluid specimens from 113 pediatric patients were tested using two rapid antigen detection assays for Streptococcus pyogenes. Results were compared to culture, Gram stain, and PCR results. Each rapid antigen assay detected 9 out of 10 (90%) PCR-positive samples, with 100% specificity. These antigen detection assays are useful to provide microbiological diagnosis of empyema caused by S. pyogenes.

High incidence of respiratory viruses in critically ill children

Background: Acute lower respiratory tract illness is a major cause of hospitalization, respiratory failure, and death in children worldwide. However, infectious agents are not identified in >50% of these clinical illnesses using traditional methods such as rapid antigen detection assays or viral culture. Objective: To conduct a pilot study using RT-PCR to determine the frequency of respiratory viruses in critically ill children with respiratory compromise. Methods: Critically ill children admitted to the Pediatric Critical Care Unit during 2 winter seasons with respiratory compromise or failure were prospectively enrolled. Respiratory tract specimens (tracheal lavage or nasal wash) were tested for 11 different viruses using realtime RT-PCR, including respiratory syncytial virus (RSV), human metapneumovirus (HMPV), influenza A (IAV) and B (IBV), parainfluenza viruses (PIV) 1-3, coronaviruses OC43, 229E, and Netherlands, and human rhinovirus (HRV). Results: Thirty-two children were enrolled, 28 (88%) of which required mechanical ventilation. Clinical diagnoses included bronchiolitis, croup, asthma, apnea, and pneumonia. Of these patients, 28/32 (88%) tested positive for one of eleven viruses by RT-PCR. Compared to antigen testing for RSV, IAV, and IBV, RT-PCR detected an additional nine infections for an increased yield of 53%. RT-PCR also detected coronavirus infections in five patients, including three patients with Netherlands coronavirus. Co-infection with more than one virus was detected in 8/32 (25%). None of the 32 patients tested positive for HMPV, PIV 1, or PIV 3. Conclusions: We detected a respiratory virus in 88% of children with respiratory failure. RT-PCR significantly increased the identification of viruses compared to antigen testing. RSV, influenza A, and coronaviruses were the most common viruses detected. Our data suggest that a viral etiology can be identified in the majority of childhood episodes of severe respiratory disease and that RT-PCR is a useful addition to the diagnostic approach.

Detection of enteroviruses in clinical samples of patients with aseptic meningitis by rapid antigen detection assay

Human enteroviruses represent the most common etiological agents of aseptic meningitis. Rapid diagnosis of aseptic meningitis affects the management of patients. The aim of this work was to assess the efficacy of rapid antigen detection (RAD) assay for the confirmation of human enteroviruses in comparison to that of the conventional cell culture (CCC), and to identify the serotypes associated with aseptic meningitis cases in the Autonomous Province of Vojvodina. RAD assay was carried out using centrifugation of inoculated Vero, RD and HEp-2 cell cultures and indirect immunofluorescence with Pan-Enterovirus 2E11 reagent (Millipore-Chemicon). CCC was performed using the same type of cells and the same immunofluorescence reagent for enterovirus confirmation. Out of 70 analyzed samples (29 cerebrospinal fluid specimens, 27 stool specimens, 9 rectal and 5 throat swabs), 36 (51.4%) were positive by immunofluorescence after CCC and 34 (48.6%) by the RAD assay. The sensitivity of RAD assay was 94.4% and specificity 100%. Detection time of enteroviruses by the RAD assay was 3 days, and by CCC varied from 3 to 13 days (mean time 6.1 days). Within 72 h from inoculation, a cytopathic effect (CPE) occurred in only 19 (47.5%) of 40 CPE positive samples. Serotyping revealed 11 types of enteroviruses: coxsackievirus A16, B3, B4, B5, and echovirus 2, 4, 6, 11, 13, 16 and 30. The RAD assay was slightly less sensitive than CCC and significantly shortened the detection time of enteroviruses, so it may be useful in rapid diagnosis of enteroviral meningitis.

Changing trends in prevalence of different Plasmodium species with dominance of Plasmodium falciparum malaria infection in Aligarh (India)

Objective To determine the prevalence of malaria in Aligarh and analyze species dominance in different years over a decade. Methods Diagnosis of malaria was done using microscopy as gold standard, rapid antigen detection assays and quantitative buffy coat (QBC) assays. Giemsa stained blood smear examination was done, thick and thin films were examined for presence of different Plasmodium spp. Rapid antigen detection assays employing detection of HRP-2 and parasite lactate dehydrogenase antigen (pLDH) by immunochromatography was done in patients whose blood smear found to be negative by conventional Giemsa slide examination. QBC was done in cases where there is strong clinical suspicion of malaria with blood smear negative, in patients with chronic malaria, splenomegaly, or in those patients who had inadequate treatment and for post-treatment follow up. Results Plasmodium vivax and Plasmodium falciparum were only species detected in our hospital. Overall prevalence of malaria in Aligarh was found to be 8.8%. The maximum prevalence of 20.1% was observed in year 2008 and lowest 2.3% in 2002. Conclusions High prevalence of malaria is observed in this part of country with dominance of both species particularly Plasmodium falciparum should be monitored and factors accounting for occurrence should be studied to employ effective control measures.

Unique Finding of a 2009 H1N1 Influenza Virus-Positive Clinical Sample Suggests Matrix Gene Sequence Variation

The 2009 H1N1 influenza virus has rapidly spread all over the world. At this time, regions in the northern hemisphere are at the beginning of the typical annual respiratory virus season, although the 2009 H1N1 virus has been highly prevalent for the last several months. This year, it is likely that the 2009 H1N1 and seasonal influenza viruses will coexist for a period of time. Rapid and accurate laboratory detection of influenza virus and its subtype is very important for selection of appropriate antiviral therapy and initiation of infection control measures for hospitalized patients. However, with limited useful assays available, this is a challenging task. While quick and easy to perform, rapid influenza virus antigen detection assays are known to suffer from low sensitivity for the 2009 H1N1 virus (1, 2). Reverse transcription-PCR (RT-PCR) is not only sensitive compared to virus culture but also much more rapid than culture and, therefore, is widely used (3). Here, we report our finding of a highly unusual case of 2009 H1N1 influenza.

Prospective Evaluation of Rapid Antigen Tests for Diagnosis of Respiratory Syncytial Virus and Human Metapneumovirus Infections

Respiratory syncytial virus (RSV) and human metapneumovirus (hMPV) are two important viral pathogens that cause respiratory tract infections in the pediatric population. The rapid detection of these agents allows the prompt isolation and treatment of infected patients. In the present prospective study, we evaluated the performances of four rapid antigen detection assays, including a rapid chromatographic immunoassay (CIA) for RSV (Directigen EZ RSV; Becton Dickinson, Sparks, MD), a direct fluorescent-antibody assay (DFA) for RSV (Bartels; Trinity Biotech, Carlsbad, CA), and two DFAs for hMPV manufactured by Diagnostic Hybrids Inc. (DHI; Athens, OH) and Imagen (Oxoid Ltd., Basingstoke, Hampshire, United Kingdom). The clinical specimens tested comprised 515 nasopharyngeal aspirates submitted to the Clinical Microbiology Laboratory at Hartford Hospital from 1 November 2006 to 21 April 2007. Compared to the results of real-time reverse transcription-PCR (RT-PCR), the CIA had a sensitivity of 79.8% and a specificity of 89.5%. The RSV DFA with Bartels reagents showed a sensitivity of 94.1% and a specificity of 96.8%. For hMPV, the sensitivity and specificity were 62.5% and 99.8%, respectively, for the DHI DFA and 63.2% and 100%, respectively, for the Imagen DFA. The hands-on and test turnaround times for CIA were 10 and 30 to 60 min, respectively, and the hands-on and test turnaround times for the RSV and hMPV DFAs were 30 and 105 min, respectively. We conclude that while the RSV CIA is user-friendly, it lacks sensitivity and specificity, especially during off-peak months. In contrast, the RSV DFA is more sensitive and specific, but interpretation of its results is subjective and it demands technical time and expertise. Similarly, both hMPV DFAs are highly specific in comparison to the results of RT-PCR, but their sensitivities await further improvements.