There have been recent FDA warnings regarding false negative molecular tests.
This is a timely reminder that false negative results have been well documented with molecular tests, especially early in the course of infection. Negative tests should always be interpreted in the context of the exposure history and clinical presentation of person being tested. See additional resources on this page including: Interpreting Viral Test Results-Key Points, Considerations for Testing Asymptomatic Persons, and the FAQ What if I am concerned about a result being a false negative?
The SARS-CoV-2 testing landscape is continually changing as new tests receive emergency use authorizations (EUAs) from the FDA. Molecular, antigen, and serology tests are now available and additional technologies are anticipated. The availability of evidence regarding the performance of these different tests as well as guidelines for their best use in different populations is still limited. This webpage provides an overview of SARS-CoV-2 tests with a focus on the use and interpretation of diagnostic (viral) tests. The LAC DPH Testing Guidelines and Resources page hosts local testing and laboratory information as well as testing recommendations.
LAC DPH is examining the emerging data, following the development of new testing technologies, and staying alert to significant changes in policies and guidance from the CDC, the California Department of Public Health (CDPH), FDA, and other public health authorities concerning COVID-19 testing. Providers are encouraged to revisit these pages which are updated frequently in light of changing test technologies, data, policies and guidance.
Only tests with emergency use authorizations (EUAs) from the FDA should be used for patient care. A wide variety of molecular tests, antigen tests, and serologic tests now have EUAs.
It is particularly important for clinicians to review the EUA for any SARS-CoV-2 test(s) they are using to understand the specific performance characteristics and the instructions for use. All SARS-CoV-2 EUAs are listed on the FDA COVID-19 Emergency Authorization website.
No tests give a 100% accurate result; tests need to be evaluated to determine their sensitivity and specificity, ideally by comparison with a gold standard. For COVID-19, there is no clear-cut gold-standard which makes the evaluation of test accuracy challenging. No clinical performance data is required for an EUA and there is limited information on how these different COVID-19 tests perform in real world settings.
Test accuracy is dependent on many factors including the test technology, specimen collection and handling, pretest probability of disease, the timing of test, and the specific characteristics of the assay. Clinicians should review the EUA and supporting materials for any SARS-CoV-2 test(s) they are using to understand the specific performance characteristics, the sample collection, storage and transport instructions), and what negative and positive test results mean.
Both molecular and antigen tests have been approved by the FDA for waived testing at the point-of-care setting. There is only one serology test approved at the current time for POC testing.
For facilities performing only waived SARS-CoV-2 tests, a temporary allowance has been made during the CA emergency. Facilities may begin waived SARS-CoV-2 testing as soon as they have submitted an application for a California Clinical Laboratory Registration and received acknowledgment of receipt. See CDPH Temporary Allowance for Waived SARS-CoV-2 Testing for more information.
All positive and non-positive (negative, indeterminate, unsatisfactory) test results from POC devices must be reported to Public Health.
It is critically important that testing sites observe best practices for handling and performing tests on infectious disease specimens and fully train all staff engaged in sampling and test operations. The CDPH LFS Branch website provides guidance on the operation of COVID-19 testing and other resources including Information for Physician Offices and Urgent Care Clinics. For additional information on POC testing see CDC Guidance for SARS-CoV-2 POC Testing.
DIAGNOSTIC (VIRAL) TESTS
Shows current infection
Amplifies specific fragment of viral RNA using nucleic acid amplification
Lateral flow immunoassays to detect viral surface proteins
Type of sample*
|Nasopharyngeal (NP), oropharyngeal, or nasal swab; saliva; lower respiratory tract specimens||NP or nasal swab.|
Authorized for use at point of care (POC)
|Most are not, but some are.||Yes.|
Test turn-around time
Standard RT-PCR: < 48 hours (but due to lab volume and capacity, may be significantly longer due to backlogs)
POC tests range from 15-45 min
|Range from 15-30 min|
Interpretation of positive tests
RNA from SARS-CoV-2 was detected. The patient is infected with the virus and considered to be contagious.
Note: A positive test in an asymptomatic person within 3 months of a prior lab-confirmed COVID-19 infection likely represents detection of non-viable virus.
|Antigens from SARS-CoV-2 were detected. The patient is infected with SARS-CoV-2 virus and considered to be contagious.|
Interpretation of negative tests
Negative tests should always be interpreted in the context of the exposure history and clinical presentation of person being tested.
A negative result in a patient with a high pre-test probability of infection (e.g. there is strong clinical suspicion for COVID-19 and/or recent exposure) has an increased likelihood of being a false negative.
A negative result in a patient with low pre-test probability of infection is more likely to be a true negative.
Negative tests should always be interpreted in the context of the exposure history and clinical presentation of person being tested.
In most cases, negative antigen tests are considered presumptive and should be confirmed with a RT-PCR test.
A negative result in a patient with a low pre-test probability of infection is more likely to be a true negative and confirmatory testing may not be needed unless important for clinical management or infection control. See CDC rapid antigen testing for more discussion.
Considerations for Use
RT-PCR is recommended for the following:
*A close contact cannot be released early from quarantine on the basis of negative viral test.
The best use of antigen tests is still being evaluated. Their intended use is in symptomatic persons early in course of infection, however they are currently being deployed and assessed for use in asymptomatic persons.
Due to their rapid turn-around time and ease of use antigen tests may be useful to rapidly rule in infected persons in the following situations:
In addition, they may be useful for the following:
*Refer to assay specific IFU for symptomatic window of detection
**A close contact cannot be released early from quarantine on the basis of negative viral test.
Molecular tests are currently the most accurate tests for diagnosing COVID-19. Molecular tests detect the unique genetic sequence of the SARS-CoV-2 virus by using nucleic acid amplification (NAA) technology such as reverse transcription polymerase chain reaction (RT-PCR) or other amplification methods (e.g. LAMP). RT-PCR is highly sensitive and specific for detecting SARS-CoV-2 virus and is the recommended “gold standard” method for diagnosing current infection. Most molecular tests are moderate or high complexity and must be run in a laboratory, however some are also authorized to be conducted in the point-of-care (POC) setting under a Clinical Laboratory Improvement Amendments (CLIA) Certificate of Waiver.
Not all molecular tests have equivalent performance. In addition to each assay’s published test performance in the EUA, the FDA is now publishing comparative performance data of molecular tests against a standard reference panel. The sensitivity of molecular tests is also dependent on the timing of testing, sampling technique, and sample type. Molecular tests are considered to be very specific.
Laboratory turn-around time (TAT) for PCR test results should be less than 48 hours, however results can be delayed if laboratories are experiencing high volumes. Currently available POC molecular tests results have a TAT ranging from 15-45 minutes.
See FDA list of EUAs for Molecular Tests for more information on the performance and use of specific authorized tests.
Antigen tests detect the presence of SARS-CoV-2 viral surface proteins. The main advantages of antigen tests are that results are available within 15-30 minutes, they are relatively simple to perform, and they are less expensive than RT-PCR tests. The currently available EUA authorized SARS-CoV-2 antigen tests are all approved as POC tests for settings with a CLIA waiver and they are intended for use on symptomatic persons within 5-12 days of symptom onset (the number of days varies by manufacturer).
While antigen tests are considered very specific for the virus, they are less sensitive than RT-PCR.
See FDA’s list of EUAs for Antigen Tests for more information about the performance and use of specific authorized tests.
See FDA letter to Healthcare Providers Potential for False Positive Results with Antigen Tests alerting providers that false positive results can occur, especially when users do not follow the instructions for use or when testing in low prevalence settings. Visit FAQs below for more on false positive results.
The best use of antigen tests is still being evaluated. While their intended use is in symptomatic persons early in the course of infection, they are often used in asymptomatic persons. Antigen tests can augment other testing efforts, especially in settings where RT-PCR testing capacity is limited or testing results are delayed. Rapid antigen tests should add both clinical and infection control value as they can rapidly “rule in” persons with SARS-CoV-2 infection, particularly when the likelihood of infection is high, such as patients with symptoms suggestive of infection, with recent close contacts to a known case, or persons that are part of an outbreak setting. Negative antigen tests in these situations should be confirmed by RT-PCR.
There is limited data to guide the use of antigen tests for screening asymptomatic persons in the absence of a known exposure. Antigen testing guidelines released by the CDC and CDPH outline scenarios where antigen tests may be considered for screening asymptomatic persons.
To learn more, see current antigen testing guidance:
Serology Tests detect waning or past SARS-CoV-2 virus infection indirectly, by measuring the antibody response to the virus. Serology assays should not replace direct viral detection methods for diagnosing an active SARS-CoV-2 infection. At this point in time, until the presence, durability, and duration of immunity is established, serology tests they should not be used to determine immune status. Healthcare providers and systems considering using serologic tests for patient care should adopt strategies to minimize false positive results as outlined in the CDC antibody testing guidance.
Use of Serologic Tests
CDC Recommendations for Use of Serologic Tests include:
Non-diagnostic uses of serological tests:
When SARS-CoV-2 serology tests should not be used:
To learn more, see current serology guidance:
Routine viral testing of asymptomatic persons in the general population in the absence of an exposure has limited value. Due to uncertainties in test performance in asymptomatic individuals, particularly with those at lower likelihood of infection, the meaning of a positive or negative test result becomes less clear.
There are situations where testing asymptomatic people is strongly recommended, such as recently exposed close contacts of a known case, persons who are part of contact investigations and/or outbreaks, and routine testing of staff and residents of nursing homes. In the absence of clear clusters of cases or known exposures, the utility of routinely testing asymptomatic persons in other situations and settings such as schools, hospitals, and workplaces is not clear. See Smart Testing for COVID-19 Virus and Antibodies, Center for Infectious Disease Research and Policy for more discussion.
Serial antigen testing of asymptomatic persons within a closed congregate setting, such as a long-term care facility or a correctional facility, has been suggested by the CDC as a potential strategy to rapidly identify cases of SARS-CoV-2 infection to prevent further transmission in the facility. CDC cites modeling evidence shows that outbreak control depends largely on the frequency of testing and the speed of reporting and is only marginally improved by high test sensitivity.
It is important to emphasize that negative test results only indicate that the test did not detect the virus at the time the test was taken. With widespread community transmission, any person who interacts with other people runs a daily risk of acquiring COVID-19. This daily risk increases in crowded places, in confined spaces (especially indoors), with close contact, and when protective actions, such as maintaining physical distancing, correctly using face coverings when around others, frequent hand hygiene, and other job-specific protective measures, are not followed.
A false negative result can happen when an infected individual is still incubating the infection (i.e., that the test was taken too early in the course of the infection) and/or that the test simply failed to detect the SARS-CoV-2 virus.
A negative test does NOT mean the person can safely ignore physical distancing and mask requirements. Refer the patient to Information about Testing.
Negative SARS-CoV-2 viral test results should always be interpreted in the context of the patient’s exposure history and the clinical presentation of person being tested.
A negative viral test result in a patient with a high pre-test probability of infection has an increased likelihood of being a false negative. If there is strong clinical suspicion for COVID-19 (compatible symptoms and/or exposure), and a patient has a negative viral test result, the patient should continue isolation and be managed as a presumed positive.
When using lower sensitivity assays (e.g. antigen tests and some molecular tests), negative results are considered presumptive and confirmation with RT-PCR is recommended, especially if important for clinical management or infection control. If the initial negative test was a RT-PCR, consider repeating the test to confirm infection.
Any patient with a positive antigen test for COVID-19 should be treated as a case and isolated.
While point of care antigen tests are reported to have a high specificity, false positives antigen tests are known to occur when instructions for use are not followed correctly or if there are inadequate quality assurance procedures. Testing sites and laboratories should follow the instructions for use and the package insert that are specific for the test system that they are using. False positives are also more likely to occur when testing persons with a low pre-test probability of infection (e.g. screening asymptomatic persons without risk factors in a low-prevalence setting).
If a false positive antigen test result is suspected, then a confirmatory RT-PCR test can be performed. Note: confirmatory RT-PCR testing is not recommended in situations where there a high pre-test probability of infection (e.g. COVID-19 is clinically suspected, they are a close contact, they live or work in a setting with an outbreak).
If confirmatory testing is pursued, the person must remain isolated until the result of the RT-PCR test is available.
Any new* positive PCR test for COVID-19 should be considered true, treated as a case, and isolated per guidelines.
RT-PCR tests are highly specific and are the current gold-standard method for diagnosing current infection with SARS-CoV-2. While false positive tests are possible with any test, due to widespread community transmission in the county, all persons with newly identified SARS-CoV-2 virus detected via RT-PCR should be considered infected and presumed contagious.
*A repeat positive PCR test within 3 months of recovery from a confirmed SARS-CoV-2 infection in the absence of symptoms likely represents persistent shedding of non-viable viral RNA. See Patients with a History of Recent Recovery from COVID-19.
When false-positive tests are suspected, CDC recommends that facilities should review and gather the following information:
A low pretest probability has a high negative predictive value but a lower positive predictive value. This means there is an increased likelihood of true negatives and false positives.
A high pretest probability has high positive predictive value and a low negative predictive value. This means there is an increased likelihood of true positives and false negatives.
For instance, if testing a population with a COVID-19 prevalence of <1% (e.g., screening asymptomatic persons in non-outbreak settings) with a single test with 99% specificity, the positive predictive value (probability that a positive test is a true-positive) could be <40%. If the prevalence is >10% (e.g., testing asymptomatic persons as part of an outbreak response) with that same test with 99% specificity, the positive predictive value may be >90%.
See CDC interpreting test results.