Rapid Diagnostic Testing for Influenza: Information for Clinical Laboratory Directors

Information for Clinical Laboratory Directors

Rapid influenza diagnostic tests (RIDTs) detect influenza viral antigens in respiratory tract specimens. Available RIDTS detect and differentiate between influenza and B viruses, but do not specifically identify or differentiate subtypes of influenza A viruses. RIDTs are not recommended for use in hospitalized patients with suspected influenza. Molecular assays, including RT-PCR, are recommended for testing respiratory tract specimens from hospitalized patients with suspected influenza because of their high sensitivity and high specificity.

Overview of Rapid Influenza Diagnostic Tests (RIDTs)

• RIDTs can provide results within approximately 15 minutes.

• Some tests are waived from requirements under the Clinical Laboratory Improvement Amendments of 1988 (CLIA) and cleared for point-of-care use.

• Most RIDTs are immunoassays that utilize antibodies against the nucleoproteins of influenza A and B viruses to detect influenza viral antigens.

• One RIDT is an immunofluorescence assay

• Some RIDTs use a digital analyzer reader device to standardize result interpretation. RIDTs that use an analyzer reader device have higher sensitivities compared to RT-PCR than RIDTs without reader devices.

• RIDTs without analyzer reader devices have low to moderate sensitivities compared to RT-PCR.

• Some RIDTs are multiplex assays that can detect antigens of influenza A and B viruses, and SARS-CoV-2.

• All RIDTs have high specificities compared to RT-PCR.

Specimen Guidelines for Rapid Influenza Diagnostic Tests (RIDTs)

RIDTs are approved for specific kinds of respiratory specimens. Specimens to be used with RIDTs should be collected as close as is possible to the start of symptoms (ideally less than four days after illness onset). In very young children, influenza viruses can be shed for longer periods; therefore, in some instances, testing for a few days after this period may still detect influenza viruses. Immunosuppressed persons may have detectable influenza viruses in respiratory specimens for prolonged periods (weeks to months).

For a list of currently available RIDTs and the respiratory specimens approved for the tests by the U.S. Food and Drug Administration (FDA), see Table 2: Rapid Influenza Diagnostic Tests (RIDTs).

Recent Developments

In 2017, the FDA re-classified RIDTs from class I to class II devices, and now requires RIDTs to meet specific minimum criteria for sensitivity and specificity. Some RIDTs are authorized for home use.

Sensitivity

Compared to RT-PCR, FDA-cleared RIDTs must achieve 80% sensitivity for detection of influenza A and influenza B viruses.

Compared to viral culture, FDA-cleared RIDTs must achieve 90% sensitivity for detection of influenza A and 80% sensitivity to detect influenza B viruses.

Specificity

Compared to RT-PCR, FDA-cleared RIDTs must achieve 95% specificity for detection of influenza A and influenza B viruses.

Compared to viral culture, FDA-cleared RIDTs must achieve 95% specificity for detection of influenza A and influenza B viruses.

Interpretation of Testing Results

Proper interpretation of testing results, particularly negative testing results is very important.

Predictive Value Depends Upon Prevalence

The positive and negative predictive values vary considerably depending upon the prevalence of influenza (level of influenza activity) in the patient population being tested.

• False-positive (and true-negative) influenza test results are more likely to occur when disease prevalence is low, which is generally at the beginning and end of the influenza season.

• False-negative (and true-positive) influenza test results are more likely to occur when disease prevalence is high, which is typically at the height of the influenza season.

Clinical Considerations of Testing When Influenza Prevalence is Low

When influenza prevalence is relatively high, the NPV is low and false-negative test results are more likely. When influenza prevalence is high, the PPV is high and positive results are more likely to be true.

The false negative rate is the number of false negatives/number of total positives, or 1-NPV.

The interpretation of negative results should take into account the clinical characteristics of the patient and the prevalence of influenza in the patient population being tested (e.g., level of influenza activity in the community). If an important clinical decision is affected by the test result and influenza is still suspected, then the RIDT result should be confirmed by a molecular assay, such as RT-PCR.

Clinical Considerations of Testing When Influenza Prevalence Is High

When influenza prevalence is relatively high, the NPV is low and false-negative test results are more likely. When influenza prevalence is high, the PPV is high and positive results are more likely to be true.

The false negative rate is the number of false negatives/number of total positives, or 1-NPV.

The interpretation of negative results should take into account the clinical characteristics of the patient and the prevalence of influenza in the patient population being tested (e.g., level of influenza activity in the community). If an important clinical decision is affected by the test result and influenza is still suspected, then the RIDT result should be confirmed by a molecular assay, such as RT-PCR.

Selecting Tests

Many factors should be considered when selecting a test, including the following:

• Tests with high sensitivity and high specificity will provide higher positive and negative predictive values, respectively.

• Tests that are CLIA-waived can be used at the point-of-care; tests that are classified as moderately complex are required to be performed in a clinical laboratory.

Information about these characteristics can be found in product inserts and scientific articles, and by contacting the manufacturers.

Changes in Recommended Procedures Can Affect Test Results

Modification by the user can affect test performances and increase false-positive and/or false-negative rates. Such modifications include:

• Using specimens for which the test is not optimized

• Using swabs that did not come with the rapid influenza diagnostic test kits [unless recommended (see package insert for specific instructions

• Improper storage or prolonged storage before specimens are tested

When Is Use of Rapid Diagnostic Tests Beneficial?

• Use of RIDTs during an outbreak of acute respiratory disease in a congregate setting can quickly help determine if influenza is the cause and to guide prompt implementation of infection prevention and control measures.

• During influenza season, testing of selected outpatients presenting with acute respiratory illnesses compatible with influenza can help establish whether influenza is present in a specific outpatient population and help health-care providers determine how to use their clinical judgment for diagnosing and treating respiratory illness. (Testing need not be done for all patients.)

• For outpatients with suspected influenza, rapid molecular assays are recommended over RIDTs because of higher sensitivities of rapid molecular assays to detect influenza A and B viruses in respiratory specimens. However, if rapid influenza molecular assays are not available, RIDTs may be helpful despite their lower sensitivities to detect influenza viruses compared with molecular assays.

• For hospitalized patients with suspected influenza, molecular assays are recommended because of their high sensitivities, and RIDTs are not recommended.

• RIDTs do not address the public health need for influenza virus isolates that can only be obtained through the collection of respiratory specimens for viral culture. Influenza virus isolates are essential for determining the match between circulating influenza virus strains and those virus strains contained in the vaccine and for aiding in the selection of new influenza vaccine virus strains.

• A positive RIDT result for influenza A virus cannot differentiate between seasonal influenza A viruses circulating among humans and influenza A viruses circulating among animals (e.g., avian influenza A viruses or swine influenza A viruses). If human infection with a novel influenza A virus of animal origin is suspected on the basis of recent exposures to poultry or pigs, the state public health department should be consulted to arrange specific RT-PCR testing for novel influenza A viruses.

Resources