Test ID: PHIV
HIV-1 Proviral DNA Qualitative Detection by PCR, Blood
Secondary ID 
A test code used for billing and in test definitions created prior to November 2011
NY State Approved Indicates the status of NY State approval and if the test is orderable for NY State clients.
Useful For Suggests clinical disorders or settings where the test may be helpful
Virologic detection of HIV-1 infection in infants of <2 years of age (an age group for which serologic tests are unreliable) born to HIV-1-infected mothers
Early detection of HIV-1 infection in children and adults prior to appearance of HIV-1 RNA, HIV-1 p24 antigen, or HIV-1 antibodies in blood
Determining eradication of HIV-1 in individuals receiving investigational highly active antiretroviral therapies
Testing Algorithm Delineates situation(s) when tests are added to the initial order. This includes reflex and additional tests.
The following algorithms are available in Special Instructions:
-HIV Testing Algorithm (excludes HIV rapid testing)
-HIV Rapid Serologic Testing Follow-up Algorithm
Special Instructions and Forms Describes specimen collection and preparation information, test algorithms, and other information pertinent to test. Also includes pertinent information and consent forms to be used when requesting a particular test
Method Name A short description of the method used to perform the test
Polymerase Chain Reaction (PCR)
(PCR is utilized pursuant to a license agreement with Roche Molecular Systems, Inc.)
Reporting Name A shorter/abbreviated version of the Published Name for a test; an abbreviated test name
Aliases Lists additional common names for a test, as an aid in searching
AIDS (Acquired Immune Deficiency Syndrome)
HIV (Human Immunodeficiency Virus)
HIV DNA Detection
HIV-1 DNA
HIV-1 Proviral
HIV-1 Qualitative
Human Immunodeficiency Virus (HIV)
Specimen Type Describes the specimen type needed for testing
Specimen Required Defines the optimal specimen. This field describes the type of specimen required to perform the test and the preferred volume to complete testing. The volume allows automated processing, fastest throughput and, when indicated, repeat or reflex testing.
Collection Container/Tube: Lavender top (EDTA)
Submission Container/Tube: Plastic vial
Specimen Volume: 2 mL
Collection Instructions:
1. Refrigerate EDTA-whole blood specimen within 6 hours of collection, and ship on ice pack in <24 hours.
2. If shipment will be delayed for >24 hours, freeze EDTA-whole blood specimen at -70 degrees C (up to 35 days) until shipment on dry ice.
Additional Information: This test is intended for diagnosis of acute HIV-1 infection, including for detection of HIV-1 infection in children <18 months of age when serologic tests are not useful (due to presence of maternal HIV antibodies).
Forms: If not ordering electronically, submit a General Request Form (Supply T239) with the specimen.
Specimen Minimum Volume Defines the amount of specimen required to perform an assay once, including instrument and container dead space. Submitting the minimum specimen volume makes it impossible to repeat the test or perform confirmatory or perform reflex testing. In some situations, a minimum specimen volume may result in a QNS (quantity not sufficient) result, requiring a second specimen to be collected.
Reject Due To Identifies specimen types and conditions that may cause the specimen to be rejected
| Hemolysis | Mild OK; Gross OK |
| Lipemia | Mild OK; Gross OK |
| Icterus | NA |
| Other | NA |
Specimen Stability Information Provides a description of the temperatures required to transport a specimen to the laboratory. Alternate acceptable temperature(s) are also included.
| Specimen Type | Temperature | Time |
|---|---|---|
| Whole Blood EDTA | Frozen (preferred) | |
| Refrigerated | 72 hours | |
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
Human immunodeficiency virus (HIV)-1 infection in humans is usually confirmed by detection of HIV-1-specific antibodies in serum. However, serologic testing may not reliably identify HIV-1 infection in neonates with passively acquired maternal HIV-1 antibodies or with incompletely developed immune systems, in individuals with early infection (<30 days from infection), or with "indeterminate" antibody profiles by Western blot assays. In these situations, detection of HIV-1 nucleic acids (RNA or proviral DNA) by PCR can provide early evidence of HIV-1 infection (approximately 10-14 days after infection), when results of routine diagnostic assays are inconclusive.
Upon entry into human cells (including peripheral blood mononuclear cells), the HIV-1 RNA is converted into complementary DNA (cDNA) by reverse transcription. These linear cDNA strands are then integrated into the host cell genome, thus representing the proviral form of HIV-1. mRNA, transcribed from the proviral DNA, is used to synthesize the proteins required to make new viral particles. These proteins and viral RNA are packaged in the host's cytoplasm and released from the cell, completing the life cycle of the virus.
The presence of integrated HIV-1 proviral DNA can be detected by a PCR that targets a segment of the highly conserved HIV-1 gag gene. Clinical studies have indicated that detection of HIV-1 proviral DNA in whole blood specimens by PCR is highly sensitive (>95%) and specific (>98%) for the presence of early HIV-1 infection in infants <2 years old.
HIV-1 virologic testing is recommended at birth, 1 to 2 months of age, and 6 months after birth, in infants born to HIV-1-infected mothers. Two serially positive HIV-1 virologic test results (HIV-1 proviral DNA or HIV-1 RNA) are necessary for the diagnosis of HIV-1 infection in infants <2 years of age.
Reference Values Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.
Negative
Interpretation Provides information to assist in interpretation of the test results
A positive result is consistent with HIV infection (see Cautions). Two serially positive HIV-1 virologic test results (HIV-1 proviral DNA or HIV-1 RNA) are necessary for the diagnosis of HIV-1 infection in infants <2 years of age.
A negative result indicates that HIV-1 DNA was not detected in the specimen (see Cautions). The lower limit of detection is 66 copies/mL.
Indeterminate results indicate that the presence or absence of HIV-1 DNA could not be determined with certainty after repeat testing of the clinical specimens in the laboratory, possibly due to PCR inhibition. Submission of a new specimen for testing is recommended.
Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances
This assay should not be used as a screening test or a primary diagnostic test for HIV-1 infection except in infants of <2 years of age born to HIV-1-infected mothers.
This assay is optimized for the detection of group M subtypes (A to H) of HIV-1, but not groups N or O.
Diagnosis of HIV-1 infection should not rely solely upon the result of an HIV-1 proviral DNA assay. A positive result should be considered in conjunction with clinical presentation and additional established diagnostic tests prior to establishing a diagnosis. A negative result indicates only the absence of HIV-1 proviral DNA in the specimen tested and does not exclude the diagnosis of disease. Negative results should be interpreted with caution, considering the patient's risk factors for HIV infection, the analytical sensitivity of the assay (95% detection limit of 66 copies/mL), and genotype of the infecting HIV-1 strain. Follow-up testing is recommended for high-risk patients with initially negative test results.
Clinical Reference Provides recommendations for further in-depth reading of a clinical nature
1. DeSimone JA, Pomerantz RJ: New methods for the detection of HIV. Clin Lab Med 2002;22:573-592
2. Working Group on Antiretroviral Therapy and Medical Management of HIV-Infected Children. Guidelines for the use of antiretroviral agents in pediatric HIV infection. Available from URL: http://www.aidsinfo.nih.gov/ContentFiles/PediatricGL_SupIPDA.pdf 2005, pp 4-5
Method Description Describes how the test is performed and provides a method-specific reference
HIV-1 Proviral DNA is extracted and purified from whole blood by the automated MagNA Pure LC instrument (Roche Applied Science, Indianapolis, IN) using the MagNA Pure LC DNA Isolation Kit-Large Volume. The specimen input volume is 500 mcL with a final elution volume of 110 mcL.
The double-stranded DNA is denatured by heat to expose the target region to labeled primers. The amplification target, a highly conserved region of the GAG genome, is bordered by the primer pair SK145 and SKCC1B. The 2 biotinylated oligonucleotide primers complementary to the amplification target sequence will bind to the target region on the DNA (native or internal control DNA). rTth polymerase links deoxynucleotide triphosphates, (dATP, dGTP, dCTP, dUTP) extending in the 5' to 3' direction to produce biotinylated complementary DNA sequences called amplicons. During PCR, controlled fluctuations in temperature allow repeated denaturation, annealing, and extension processes resulting in a geometric increase in the target sequences. DNA copies from previous cycles become templates in subsequent amplification periods. The resulting amplicon is 155 bp in length. HIV-1 internal control is added to identify clinical specimens containing inhibitory substances or test failure due to inadequate individual specimen processing. The internal control is introduced into each specimen during the extraction steps to serve as an extraction and amplification control for each independently processed specimen.
Uracil-N-glycosylase (UNG) is used to prevent contamination from previous PCR reactions. UNG will excise any dUTP found in previously amplified DNA. (Naturally occurring DNA will contain dTTP.) Before the PCR reaction is initiated, the thermal cycler is set at 55 degrees C to optimally activate UNG for 2 minutes. After the amplification cycles are completed, the UNG is inactivated by a denaturing solution containing sodium hydroxide.
The amplified DNA is incubated in polystyrene wells containing immobilized bovine serum albumin-conjugated probe (SK 102) that is specific to the biotinylated amplicons. After incubating for 1 hour, the unbound reactants are washed away. An avidin-horseradish peroxidase conjugate is added and incubated. Unbound reactants are washed away. A substrate solution containing hydrogen peroxide and 3,3',5,5'-tetramethylbenzidine (TMB) is added. In the presence of hydrogen peroxide, the bound horseradish peroxidase catalyzes the oxidation of TMB to form a color complex. The reaction is stopped by the addition of a weak acid and the absorbance is read at 450 nm. A fixed optical density cutoff is used to determine whether a specimen is positive or negative.(Package insert: Amplicor HIV-1 DNA Test, version 1.5, Roche Diagnostics, Indianapolis, IN, March 2005)
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
Varies
Analytic Time Defines the amount of time it takes the laboratory to setup and perform the test. This is defined in number of days. The shortest interval of time expressed is "same day/1 day," which means the results may be available the same day that the sample is received in the testing laboratory. One day means results are available 1 day after the sample is received in the laboratory.
Maximum Laboratory Time Defines the maximum time from specimen receipt at Mayo Medical Laboratories until the release of the test result
Specimen Retention Time Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded
Performing Laboratory Location The location of the laboratory that performs the test
Test Classification Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer's instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR), Investigation Use Only (IUO) product, or a Research Use Only (RUO) product.
CPT Code Information Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Medical Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.
87535
LOINC® Code Information Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the result codes returned for this test or profile.
| Result ID | Reporting Name | LOINC Code |
|---|---|---|
| 88635 | HIV-1 Proviral DNA, B | 48023-6 |
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