Histoplasma DNA detection by PCR

General Information

Lab Name

Lab Code

HISDNA

Epic Ordering

Histoplasma DNA detection by PCR

Description

Detection of Histoplasma capsulatum DNA.

Histoplasma capsulatum (or Ajellomyces capsulatum) causes Histoplasmosis which mainly affects lungs. Infected individuals generally exhibit no clinical manifestations or ill effects. Acute phase is manifested by non-specific respiratory symptoms that are cough or flu like. Chronic Histoplasmosis can manifest tuberculosis like symptoms and can also lead to disseminated disease that can be fatal if not treated. Diagnosis by detection of antigen (EIA) and detection of antibody are widely used. Culture based confirmatory diagnosis is time consuming due to slow growth and manifestation of specific phenotype. PCR methods have been recently used to confirm presence of Histoplasma capsulatum in biopsy materials.

Histoplasma PCR detects Histoplasma capsulatum (or Ajellomyces capsulatum) with higher specificity and sensitivity than the broad-range fungal PCR. The analytical sensitivity is 1 genome per PCR reaction.

Taxonomic identifications are reported based on sequencing of the amplification product.

For more information on this test, to download a requisition form, and for a list of acceptable and unacceptable specimens, please refer to: Molecular Diagnosis Microbiology Section Website

References

Kauffman et al. (2007) Histoplasmosis: Clinical and Laboratory Update in Clinical Microbiology Reviews p. 115-132

//en.wikipedia.org/wiki/Histoplasmosis

Synonyms

Fungal sequencing, Histoplasma detection, Histoplasma identification, Histoplasma PCR, Histoplasma sequencing, Histoplasmosis PCR, molecular Histoplasma, universal PCR

Components

Code	Name
HISRS	Histoplasma PCR: Detection, nested ITS
HISID	Histoplasma PCR: Identification, nested ITS
HISSI	Histoplasma PCR: Specimen Description
HISSPI	Histoplasma PCR: External Identifier
HISSR	Histoplasma PCR: Special Requests
HISSC	Histoplasma PCR: Specimen Comments
HISNAE	Histoplasma PCR: Specimen DNA Extraction
HISREV	Histoplasma PCR: Pathologist Review
HISME	Histoplasma PCR: Method Note

Interpretation

Method

DNA extraction, nucleic acid purification, polymerase chain reaction (PCR), sequencing

Reference Range

See individual components

References

Kauffman et al. (2007) Histoplasmosis: Clinical and Laboratory Update in Clinical Microbiology Reviews p. 115-132

//en.wikipedia.org/wiki/Histoplasmosis

Guidelines

Ordering & Collection

Specimen Type

Tissue (Fresh frozen or paraffin-embedded), Fluid (see Acceptable Specimens for details)

Collection

Acceptable specimens are listed below. Please see this page for complete specimen collection and handling instructions.

Specimens should be collected into a DNA free container labeled with at least two identifiers.

Acceptable Specimens

Fresh frozen tissue
Fresh frozen fluid: any body fluid is acceptable if it is not listed under Unacceptable Specimens. See common examples below.
Formalin Fixed Paraffin-embedded tissues (FFPE, PET): blocks, scrolls, and unstained slides are acceptable
eSwabs and UTM (universal transport media)

Common acceptable body fluid examples: cerebrospinal fluid, pleural fluid, pericardial fluid, urine, bronchial lavage, joint fluid, bone marrow, vitreous fluid, etc.

Additional Acceptable Specimens

Sputum

Unacceptable Specimens

Blood, serum, plasma, stool
No citrated or heparinized solutions
No SPS (Sodium Polyanethole Sulfonate) e.g. Wampole Isolator Tubes
Tissues floating in formalin
Swab/fluid collected in tube containing agar

Handling Instructions

Fresh frozen tissue/fluid specimens should be submitted and maintained on dry ice.

Formalin Fixed Paraffin-embedded tissues (FFPE, PET) can be sent ambient or with ice packs during warmer summer months to prevent melting.

Optimal Quantity:

Fresh Tissue: 0.3-1.0 cm^3
Fluid: 0.2-1 mL
Formalin Fixed Paraffin-embedded Tissue (FFPE/PET): blocks are preferred and will be sent back to client upon completion of testing
Scrolls/unstained slides: cross-sectional area >1cm^2 send 10 sections of 10µm thickness, if <1cm^2 send 20 sections if available

Please note: We do not need a separate specimen aliquot for each test ordered. Only a single specimen aliquot or block of optimal quantity is necessary for performing multiple tests. If multiple aliquots or blocks of optimal quantity are sent, up to 2 will be pooled.

Quantity

requested: See Optimal Quantity above
minimum: Specimens below optimal quantity are acceptable for testing, however, diagnostic yield is generally proportional to specimen size.

Processing

Processing: UWMC/HMC – store and send specimen refrigerated and freeze specimens at -20°C upon arrival in UW-ML Microbiology. If specimen storage and transport will exceed 8 hours, freeze at -20°C.

Freeze specimens at -20°C upon arrival

Outside Laboratories: Freeze specimens in sterile containers at -20°C. Transport all specimens on dry ice.

Performance

LIS Dept Code

Micro Molecular Diag (MMD)

Performing Location(s)

UW-MT	Microbiology, Molecular Diagnostics 206-520-4600 Clinical Lab, Room # NW177, University of Washington Medical Center, 1959 NE Pacific street, Seattle, WA 98195

Frequency

Fresh frozen tissues/fluids result in 2-3 business days after receipt of specimen. Formalin Fixed Paraffin-embedded tissues result in 3-4 business days after receipt of specimen.

Available STAT?

Billing & Coding

CPT codes: 87801
LOINC: 5015-3
Interfaced Order Code: UOW4424