Acanthamoeba and Balamuthia species PCR

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General Information

Lab Name
Acanthamoeba and Balamuthia species PCR
Lab Code
AMBPCR
Epic Ordering
Acanthamoeba and Balamuthia species PCR
Description

Detection and identification of Acanthamoeba and Balamuthia species DNA.

Free living amoeba of the Acanthamoeba and Balamuthia families are opportunistic protozoans that are widely distributed in the environment and are well recognized to produce serious human infections, including cutaneous infections, keratitis, and encephalitis. Although these infections are relatively rare, they can incur significant morbidity and mortality.

PCR has emerged as a key tool in diagnosing disease caused by these organisms, and has been shown to outperform conventional tests in terms of turnaround times, sensitivity, and specificity (1,2).

Our broad range PCR assay offers highly sensitive molecular identification of Acanthamoeba and Balamuthia species, having a limit of detection of a single organism per reaction. Uniquely, our broad-range test provides clinical diagnosis of infections caused by either of those genera using a single test.

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

1. K. Khairnar, G.S. Tamber, F. Ralevski, D.R. Pillai Comparison of molecular diagnostic methods for the detection of Acanthamoeba spp. from clinical specimens submitted for keratitis.
Diagn Microbiol Infect Dis, 70 (2011), pp. 499-506

Synonyms
18S PCR, Acanthamoeba PCR, Amoeba PCR, Balamuthia PCR, parasite identification, protozoan identification
Components

Interpretation

Method

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

Reference Range
See individual components
References

1. K. Khairnar, G.S. Tamber, F. Ralevski, D.R. Pillai Comparison of molecular diagnostic methods for the detection of Acanthamoeba spp. from clinical specimens submitted for keratitis.
Diagn Microbiol Infect Dis, 70 (2011), pp. 499-506

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 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?
No

Billing & Coding

CPT codes
87801
LOINC
18725-2
Interfaced Order Code
UOW4211