HIV RNA Detection by RT_PCR (Qualitative)

Immunity

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Detect infection with Human Immunodeficiency Virus

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HIV RNA Detection by RT-PCR (Qualitative)

Why is it done?
- Detects the presence or absence of HIV RNA (ribonucleic acid) in blood plasma using Reverse Transcription-Polymerase Chain Reaction (RT-PCR) technology
- Confirms HIV infection in patients with positive screening tests (antibody or antigen tests)
- Detects HIV during the window period (acute infection before antibodies develop), typically 7-28 days after exposure
- Screens blood and organ donors to prevent transmission through transfusion or transplantation
- Evaluates occupational exposures (needlestick injuries) and post-exposure prophylaxis (PEP) effectiveness
- Assesses vertical transmission risk in pregnant women to determine prevention of mother-to-child transmission (PMTCT) strategies
- Assists in differential diagnosis of acute retroviral syndrome (acute HIV infection presenting with flu-like symptoms)
Normal Range
- Result Interpretation: This is a qualitative test with binary (yes/no) results rather than a quantitative measurement
- Negative/Normal Result: HIV RNA not detected (reported as 'Not Detected' or 'Negative')
- Positive/Abnormal Result: HIV RNA detected (reported as 'Detected' or 'Positive')
- Detection Threshold: Typically 20-50 copies/mL depending on the specific assay used; modern assays can detect as low as 1-5 copies/mL
- Units of Measurement: Not applicable for qualitative testing; results are reported as detected or not detected
- Indeterminate/Inconclusive Result: May occur due to technical issues; repeat testing is recommended after 1-2 weeks
Interpretation
- HIV RNA Detected (Positive Result):
  - Indicates active HIV infection with viral replication
  - Patient is potentially infectious and capable of transmitting HIV to others
  - Establishes definitive diagnosis of HIV infection when paired with positive confirmatory antibody test
  - Critical in acute infection phase before antibodies are detectable
- HIV RNA Not Detected (Negative Result):
  - Most likely indicates absence of HIV infection (if obtained >2-3 weeks post-exposure)
  - In known HIV-positive patients: indicates undetectable viral load (therapeutic goal with antiretroviral therapy), meaning no clinical disease progression and negligible transmission risk ('Undetectable = Untransmittable')
  - If drawn <2 weeks post-exposure, may represent window period; repeat testing recommended
  - Safe for blood/organ donation
- Factors Affecting Test Interpretation:
  - Timing of test: Earlier detection possible with NAT (nucleic acid testing) than antibody tests
  - HIV viral load variations: may fluctuate even with undetectable levels, especially if treatment adherence is suboptimal
  - Specimen quality and storage conditions affect sensitivity
  - HIV drug resistance mutations may occasionally affect assay performance
  - Different HIV subtypes and variants may have varying detection rates
- Clinical Significance:
  - Gold standard confirmatory test for HIV diagnosis in most cases
  - Helps distinguish between acute infection, chronic infection, and treatment success
  - Essential for determining public health intervention strategies and transmission risk
Associated Organs
- Primary Organ Systems Affected by HIV:
  - Immune system: Progressive destruction of CD4+ T lymphocytes (helper T cells) leading to immunodeficiency
  - Lymph nodes and lymphoid tissues: Site of persistent viral replication and immune dysfunction
  - Central nervous system: Risk of HIV-related dementia and opportunistic infections (cryptococcal meningitis, toxoplasma encephalitis)
  - Gastrointestinal tract: Source of opportunistic infections and increased permeability leading to bacterial translocation
  - Lungs: Increased risk of Pneumocystis jirovecii pneumonia (PCP) and tuberculosis
- Conditions Associated with HIV Infection:
  - AIDS-related conditions: Opportunistic infections (CMV, MAC, candidiasis, cryptosporidiosis) and malignancies (Kaposi's sarcoma, NHL)
  - Acute retroviral syndrome: Acute HIV infection presenting with fever, fatigue, lymphadenopathy, and rash within 2-4 weeks of exposure
  - Chronic HIV disease: Progressive CD4 decline if untreated, leading to clinical AIDS
  - HIV-associated cardiovascular disease: Increased risk of myocardial infarction and stroke
  - HIV-associated kidney disease: Direct viral infection of renal cells or immune-mediated glomerulonephritis
  - HIV-associated malignancies: Increased incidence of lymphomas, cervical cancer, and anal cancer
- Potential Complications and Risks:
  - Severe immunosuppression (CD4 <50 cells/μL): Risk of disseminated infections and opportunistic cancers
  - Immune reconstitution inflammatory syndrome (IRIS): Paradoxical worsening of symptoms after starting antiretroviral therapy
  - Antiretroviral therapy (ART) side effects: Ranging from mild to serious depending on regimen
  - Drug resistance: Development of resistant viral strains with poor treatment adherence
Follow-up Tests
- Confirmatory Tests After Positive HIV RNA:
  - HIV Antibody Test (4th generation antigen/antibody test): Confirms HIV infection and rules out false positive RNA results
  - Western Blot or HIV-1/HIV-2 Differentiation Immunoassay: Confirmatory test to differentiate between HIV-1 and HIV-2 if required
- Initial Assessment Tests for Newly Diagnosed HIV Patients:
  - CD4 Count: Determines immune status and need for opportunistic infection prophylaxis; baseline assessment before starting ART
  - HIV Viral Load (Quantitative RT-PCR): Assesses degree of viral replication and baseline for treatment response monitoring
  - HIV Resistance Genotyping: Identifies drug resistance mutations to guide appropriate ART selection
  - Complete Blood Count (CBC): Evaluates for cytopenias and baseline for monitoring ART side effects
  - Liver Function Tests (LFTs): Assess hepatic function and baseline for monitoring drug toxicity
  - Renal Function Tests (Creatinine, eGFR): Determines kidney function and appropriate drug dosing
  - Lipid Panel: Assesses cardiovascular risk before starting ART
  - Hepatitis B and C Serology: Screens for coinfection requiring modified treatment approach
  - Tuberculosis Screening (TST or IGRA): Identifies active or latent TB requiring treatment
  - Opportunistic Infection Screening: CMV serology, toxoplasmosis serology if CD4 <100 cells/μL
- Ongoing Monitoring Tests:
  - HIV Viral Load (Quantitative): Every 3-6 months after starting treatment; goal is <50 copies/mL
  - CD4 Count: Every 3-6 months to monitor immune recovery
  - Liver and Kidney Function: Annually or more frequently if abnormal
  - Lipid Panel: Annually and before lipid-lowering therapy
  - HIV Resistance Testing: If viral load remains >200 copies/mL despite adherence to guide regimen changes
- Testing After Negative Result:
  - Post-exposure prophylaxis (PEP) completers: Follow-up RNA test at 4-6 weeks post-PEP if baseline rapid test was negative
  - Early test results: If test done <2 weeks post-exposure, follow-up RNA test recommended at 2-4 weeks
  - Symptomatic patients: If acute retroviral syndrome is suspected, repeat testing at 1-2 weeks if initial test negative
Fasting Required?
- Fasting Requirement: No
- Food and Drink: Patient may eat and drink normally before testing
- Medications: Continue all regular medications as scheduled; no medication restrictions for this test
- Special Instructions:
  - Bring valid photo identification and insurance card
  - Specimen collection: Venous blood sample (typically 5-10 mL) collected in EDTA or citrate tube
  - Specimen handling: Blood sample should be kept at room temperature or refrigerated and processed within 4-6 hours for optimal RNA preservation
  - Inform healthcare provider of recent vaccinations or transfusions (may not affect RNA test but important for documentation)
  - Confidentiality: Testing location must maintain strict confidentiality and follow local HIV testing regulations
  - Testing timing: Can be performed any time of day; no specific time-of-day considerations
  - Results timing: Typically available within 24-48 hours, though some rapid qualitative assays may provide results within 2-24 hours

How our test process works!