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Test Code LAB9663 Streptococcus pneumoniae PCR (Semi-Quantitative)

Performing Section

Microbiology

Purpose/Principle

In this assay, DNA extracted from patient samples is subjected to real-time PCR amplification using primers and fluorescent probes directed against three target sequences in the Spn genome: lytA, piaB, and SP2020 (5-7). The product is detected on an Applied Biosystems 7500 Taqman™ instrument using 5’-nuclease methodology and quantitated by extrapolation to a standard curve of log10 dilutions of commercially available quantified Spn whole organism. The assay is linear across five orders of magnitude and can detect as few as 100 copies/mL. An exogenous positive control with its own set of primers and probes is co-amplified in each reaction to determine extraction efficiency and detect inhibition. Results are reported as number of copies of Spn per mL of fluid sample.

Special Collection Notes

Collect aseptically according to hospital protocol.

Specimen Type

Bronchoalveolar lavage, Tracheal aspirate: Sterile Screw Capped Container

 

Pleural Fluid: Sterile Screw Capped Container

Specimen Volume

Preferred: 1.0 mL
Minimum:0.5 mL

Processing Instructions

Store and send at ambient via courier.

Temperature

Ambient

Test Components

STREPTOCOCCUS PNEUMONIAE PCR RESULT
STREPTOCOCCUS PNEUMONIAE PCR SEMI-QUANT VALUE

Reference Range

STREPTOCOCCUS PNEUMONIAE PCR RESULT

No Streptococcus pneumoniae detected

 

STREPTOCOCCUS PNEUMONIAE PCR SEMI-QUANT

No Streptococcus pneumoniae detected

TAT

72 hours

Additional Notes

*Sources requiring approval (unvalidated)
I. Sputum, 1 mL (0.5 mL minimum).
II. Tissue: 1mm3
fragment, fresh or frozen. (Requires manual extraction.)
A. Frozen tissue should be kept on dry ice or stored at -70°C until processed.
B. Fresh tissue should be stored at 2-8°C in physiological saline or a suitable transport medium
and processed as soon as possible.
C. FFPE (Formalin Fixed Paraffin Embedded); this source is less desirable due to lowered
sensitivity related to formalin fixation of tissue.

*Not Acceptable:
Blood
Urine
Upper Respiratory Sources: Sample type is acceptable for research use only. These sources are unacceptable for clinical use because this assay has not been evaluated for distinguishing between colonization and disease-causing
– Nasopharyngeal swab
– Mid-turbinate swab
– Nasal aspirate

Aliases

SPN PCR
S. pneumoniae PCR

CPT

87799