US CDC Real-Time Reverse Transcription PCR Panel for Detection of Severe Acute Respiratory Syndrome Coronavirus 2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was identified as the etiologic agent associated with coronavirus disease, which emerged in late 2019. In response, we developed a diagnostic panel consisting of 3 real-time reverse transcription PCR assays targeting the nucleocapsid gene and evaluated use of these assays for detecting SARS-CoV-2 infection. All assays demonstrated a linear dynamic range of 8 orders of magnitude and an analytical limit of detection of 5 copies/reaction of quantified RNA transcripts and 1 x 10-1.5 50% tissue culture infectious dose/mL of cell-cultured SARS-CoV-2. All assays performed comparably with nasopharyngeal and oropharyngeal secretions, serum, and fecal specimens spiked with cultured virus. We obtained no false-positive amplifications with other human coronaviruses or common respiratory pathogens. Results from all 3 assays were highly correlated during clinical specimen testing. On February 4, 2020, the Food and Drug Administration issued an Emergency Use Authorization to enable emergency use of this panel.

Use of Influenza Risk Assessment Tool for Prepandemic Preparedness.

In 2010, the Centers for Disease Control and Prevention began to develop an Influenza Risk Assessment Tool (IRAT) to methodically capture and assess information relating to influenza A viruses not currently circulating among humans. The IRAT uses a multiattribute, additive model to generate a summary risk score for each virus. Although the IRAT is not intended to predict the next pandemic influenza A virus, it has provided input into prepandemic preparedness decisions.

Comparative analytical evaluation of the respiratory TaqMan Array Card with real-time PCR and commercial multi-pathogen assays.

In this study, a multicenter evaluation of the Life Technologies TaqMan(®) Array Card (TAC) with 21 custom viral and bacterial respiratory assays was performed on the Applied Biosystems ViiA™ 7 Real-Time PCR System. The goal of the study was to demonstrate the analytical performance of this platform when compared to identical individual pathogen specific laboratory developed tests (LDTs) designed at the Centers for Disease Control and Prevention (CDC), equivalent LDTs provided by state public health laboratories, or to three different commercial multi-respiratory panels. CDC and Association of Public Health Laboratories (APHL) LDTs had similar analytical sensitivities for viral pathogens, while several of the bacterial pathogen APHL LDTs demonstrated sensitivities one log higher than the corresponding CDC LDT. When compared to CDC LDTs, TAC assays were generally one to two logs less sensitive depending on the site performing the analysis. Finally, TAC assays were generally more sensitive than their counterparts in three different commercial multi-respiratory panels. TAC technology allows users to spot customized assays and design TAC layout, simplify assay setup, conserve specimen, dramatically reduce contamination potential, and as demonstrated in this study, analyze multiple samples in parallel with good reproducibility between instruments and operators.

Development of Framework for Assessing Influenza Virus Pandemic Risk.

Although predicting which influenza virus subtype will cause the next pandemic is not yet possible, public health authorities must continually assess the pandemic risk associated with animal influenza viruses, particularly those that have caused infections in humans, and determine what resources should be dedicated to mitigating that risk. To accomplish this goal, a risk assessment framework was created in collaboration with an international group of influenza experts. Compared with the previously used approach, this framework, named the Influenza Risk Assessment Tool, provides a systematic and transparent approach for assessing and comparing threats posed primarily by avian and swine influenza viruses. This tool will be useful to the international influenza community and will remain flexible and responsive to changing information.

Pandemic preparedness and the Influenza Risk Assessment Tool (IRAT).

Influenza infections have resulted in millions of deaths and untold millions of illnesses throughout history. Influenza vaccines are the cornerstone of influenza prevention and control. Recommendations are made by the World Health Organization (WHO) 6-9 months in advance of the influenza season regarding what changes, if any, should be made in the formulation of seasonal influenza vaccines. This allows time to manufacture, test, distribute, and administer vaccine prior to the beginning of the influenza season. At the same time experts also consider which viruses not currently circulating in the human population, but with pandemic potential, pose the greatest risk to public health. Experts may conclude that one or more of these viruses are of enough concern to warrant development of a high-growth reassortant candidate vaccine virus. Subsequently, national authorities may determine that a vaccine should be manufactured, tested in clinical trials, and even stockpiled in some circumstances. The Influenza Risk Assessment Tool (IRAT) was created in an effort to develop a standardized set of elements that could be applied for decision making when evaluating pre-pandemic viruses. The tool is a simple, additive model, based on multi-attribute decision analysis . The ultimate goal is to identify an appropriate candidate vaccine virus and prepare a human vaccine before the virus adapts to infect and efficiently transmit in susceptible human populations. This pre-pandemic preparation allows production of vaccine-a strategy that could save lives and mitigate illness during a pandemic.

Development of an influenza virologic risk assessment tool.

Influenza pandemics pose a continuous risk to human and animal health and may engender food security issues worldwide. As novel influenza A virus infections in humans are identified, pandemic preparedness strategies necessarily involve decisions regarding which viruses should be included for further studies and mitigation efforts. Resource and time limitations dictate that viruses determined to pose the greatest risk to public or animal health should be selected for further research to fill information gaps and, potentially, for development of vaccine candidates that could be put in libraries, manufactured and stockpiled, or even administered to protect susceptible populations of animals or people. A need exists to apply an objective, science-based risk assessment to the process of evaluating influenza viruses. During the past year, the Centers for Disease Control and Prevention began developing a tool to evaluate influenza A viruses that are not circulating in the human population but pose a pandemic risk. The objective is to offer a standardized set of considerations to be applied when evaluating prepandemic viruses. The tool under consideration is a simple, additive model, based on multiattribute decision analysis. The model includes elements that address the properties of the virus itself and population attributes, considers both veterinary and human findings, and integrates both laboratory and field observations. Additionally, each element is assigned a weight such that all elements are not considered of equal importance within the model.

Validation of a method for preparing influenza H5N1 simulated samples.

Avian influenza virus type A subtype H5N1 and potentially other novel influenza A viruses continue to pose a concern with mutation into a form easily transmitted between humans. The ability to rapidly detect and characterize influenza viruses, and distinguish seasonal and novel influenza A viruses such as H5N1, remains important to minimize morbidity and mortality in humans. As with other rare and emerging viral pathogens, clinical specimens from persons with H5N1 infections are extremely rare. Consequently, development of standardized methods and accepted criteria are necessary for both ensuring the validity of available diagnostic methods and for assessing the potential of new diagnostic tests that can detect and differentiate H5N1 and other novel influenza A viruses. Additionally, genotypic and antigenic evolution of H5N1 poses a challenge with maintaining updated reference virus strains. In this report, a method for preparing simulated samples using defined procedures and carefully selected H5N1 virus strains is described, and the reliability for using these samples in an evaluation protocol with a laboratory test for differentiating H5N1 virus from other influenza A viruses is evaluated.