What laboratory tests are used for Human Immunodeficiency Virus (HIV) diagnosis?

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Last updated: February 19, 2025View editorial policy

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From the Guidelines

HIV Diagnosis Laboratory Tests

HIV diagnosis is typically made using a combination of tests, starting with a repeatedly reactive immunoassay followed by confirmatory Western blot or immunofluorescent assay 1.

  • The initial test is usually a fourth-generation immunoassay that detects both HIV antibodies and p24 antigens.
  • If the initial test is reactive, it is followed by a nucleic acid test (NAT) or a rapid HIV test to confirm the diagnosis 1.
  • Western blot or indirect immunofluorescence assay can also be used as supplemental tests to confirm the diagnosis 1.
  • Additionally, viral load tests, such as PCR or RNA assays, can be used to monitor the disease progression and treatment response.
  • Rapid HIV testing may use either blood or oral fluid specimens and can provide results in 5 to 40 minutes 1.
  • The U.S. Food and Drug Administration–approved tests for detection and confirmation of HIV infection include combination tests (for p24 antigen and HIV antibodies) and qualitative HIV-1 RNA 1.

From the Research

Laboratory Tests for HIV Diagnosis

The following laboratory tests are used for Human Immunodeficiency Virus (HIV) diagnosis:

  • Fourth-generation assays that simultaneously detect HIV p24 antigen and antibodies 2, 3, 4, 5, 6
  • Combination assays with separate results for anti-HIV antibodies and p24 antigen 2, 3
  • HIV nucleic acid assays 3
  • Western blot (WB) 3
  • Viral load testing 3, 5, 6
  • Dedicated HIV p24 antigen test 4
  • CD4 count monitoring to estimate the stage of infection and suggest a therapeutic approach 6

Characteristics of Fourth-Generation Assays

  • Reduce the diagnostic window between infection and diagnosis 2, 4
  • Improve detection of acute HIV infection 3, 5
  • Have high sensitivity and specificity 3
  • Can detect diverse HIV subtypes 5
  • Have similar breadth of reactivity and diagnostic utility across different commercially available assays 5

Other Diagnostic Methods

  • Biosensors are a future perspective in HIV diagnostics 6
  • Identification and detection of viral capsid p24 antigen levels is an important method in diagnostics 6

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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