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HelpTest Code KKRP Kingella kingae, Molecular Detection, PCR, Varies
Secondary ID
65201Useful For
Aiding in the diagnosis of Kingella kingae infection using tissue or synovial fluid specimens
Method Name
Real-Time Polymerase Chain Reaction (PCR)
Reporting Name
Kingella kingae PCRSpecimen Type
VariesNecessary Information
Specimen source is required.
Specimen Required
The high sensitivity of amplification by polymerase chain reaction requires the specimen to be processed in an environment in which contamination of the specimen by Kingella kingae DNA is unlikely.
Submit only 1 of the following specimens:
Specimen Type: Synovial fluid
Preferred: Lavender top (EDTA)
Acceptable: Pink top (EDTA), royal blue top (EDTA), sterile vial containing EDTA-derived aliquot, red clot tube (no anticoagulant), or sterile container
Specimen Volume: 0.5 mL
Collection Instructions: Send specimen in original tube (preferred).
Specimen Stability Information: Refrigerated (preferred) <7 days /Frozen <7 days
Specimen Type: Fresh tissue or biopsy
Sources: Bone, joint, synovium, heart valve, aorta, or endocardium
Container/Tube: Sterile container
Specimen Volume: Entire collection or 5 mm(3)- approximately the size of a pencil eraser
Collection Instructions:
1. Collect fresh tissue specimen.
2. Submit tissue only, do not add fluid to tissue
3. Refrigerate or freeze specimen.
Specimen Stability Information: Refrigerated (preferred) <7 days/ Frozen <7 days
Preferred Paraffin-embedded tissue block:
Specimen Type: Formalin-fixed, paraffin-embedded tissue block (FFPE)
Sources: Bone, joint, synovium, heart valve, aorta, or endocardium
Supplies: Tissue Block Container (T553)
Container/Tube: Tissue block
Collection Instructions: Submit a formalin-fixed, paraffin-embedded tissue block to be cut and returned.
Specimen Stability Information: Ambient (preferred)/Refrigerated
Acceptable Paraffin-embedded tissue block:
Specimen Type: Formalin-fixed, paraffin-embedded tissue block (FFPE)
Sources: Bone, joint, synovium, heart valve, aorta, or endocardium
Container/Tube: Sterile container for each individual cut section (scroll).
Collection Instructions: Perform microtomy and prepare five separate 10-micron sections. Each section (scroll) must be placed in a separate sterile container for submission.
Specimen Stability Information: Ambient (preferred)/Refrigerated
Specimen Minimum Volume
Fluid/fresh tissue or biopsy: See Specimen Required
Paraffin-embedded tissue block: Two 10-micron sections
Specimen Stability Information
| Specimen Type | Temperature | Time | Special Container |
|---|---|---|---|
| Varies | Varies | ||
Reject Due To
| Tissue in formalin, formaldehyde, or acetone Decalcified bone Bone marrow Slides |
Reject |
Clinical Information
Kingella kingae is a fastidious short gram-negative bacillus that may colonize the oropharynx of young children. Colonization may occasionally lead to invasive disease via hematogenous dissemination, primarily in children younger than 4 years of age. This most commonly results in bone and joint infection; K kingae is the most frequent cause of osteomyelitis and septic arthritis in children aged 6 to 36 months. K kingae may also cause endocarditis, involving both native and prosthetic valves, in patients of any age and is considered part of the HACEK (Haemophilus species, Aggregatibacter species, Cardiobacterium hominis, Eikenella corrodens, and Kingella species) group of organisms, known for causing culture-negative endocarditis. K kingae produces a repeat-in-toxin (RTX) toxin.
Diagnosis of K kingae infection may be challenging due to the fastidious nature of the organism in culture. Evaluation of cardiac, bone, joint tissue, or fluid by polymerase chain reaction is a useful tool for the diagnosis of some cases of K kingae infection.
Reference Values
Not applicable
Interpretation
A positive result indicates the presence of Kingella kingae DNA.
A negative result indicates the absence of detectable K kingae DNA but does not negate the presence of the organism and may occur due to inhibition of PCR, sequence variability underlying primers or probes, or the presence of K kingae DNA in quantities less than the limit of detection of the assay.
Cautions
Test results should be used as an aid in diagnosis. The single assay should not be used as the only criteria to form a clinical conclusion, but results should be correlated with patient symptoms and clinical presentation. A negative result does not negate the presence of the organism or active disease.
This assay does not detect species of Kingella other than kingae or negevensis (see Supportive Data).
This assay cross-reacts with Kingella negevensis.(1)
Supportive Data
This assay was validated by testing 30-spiked positive samples and 10-negative samples for each accepted sample type; fresh tissue, formalin-fixed paraffin-embedded tissue (FFPE), synovial fluid, and EDTA blood. No PCR inhibition was encountered. The assay was 100% sensitive and specific. The assay showed no cross-reactivity when tested with a panel of 67 bacterial isolates, including Kingella species other than kingae. The limit of detection (LOD) in fresh tissue and FFPE was 73.7 CFU/mcL. The LOD of synovial fluid was 1.3 CFU/mcL.
Clinical Reference
1. El Houmami N, Bzdreng J, Durand GA, et al: Molecular tests that target the RTX locus do not distinguish between Kingella kingae and the recently described Kingella negevensis species. J Clin Microbiol. 2017 Oct;55(10):3113-3122
2. Murphy TF: Moraxella catarrhalis, Kingella, and other gram-negative cocci. In: Bennett JE. Dolin R, Blaser MJ, eds. Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases. 9th ed. Elsevier; 2020:chap 213
3. Yagupsky P: Kingella kingae: carriage, transmission, and disease. Clin Microbiol Rev. 2015 Jan;28(1):54-79
4. Madigan T, Cunningham SA, Ramanan P, et al: Real-time PCR assay for detection of Kingella kingae in children. J Pediatr Infect Dis. 2018;13(3):216-233. doi: 10.1055/s-0038-1641603
Method Description
Nucleic acid is extracted from the specimen using the automated MagNA Pure instrument. Target specific primers are used to amplify the rxtB gene region of Kingella kingae; amplification is monitored by detecting fluorescence produced by target specific fluorescence resonance energy transfer hybridization probes. This real-time polymerase chain reaction (PCR) takes place on a LightCycler instrument. Detection of the K kingae target is performed through melting curve analysis using the LightCycler software.(Cockerill FR, Uhl JR: Applications and challenges of real-time PCR for the clinical microbiology laboratory. In: Reischl U, Wittwer C, Cockerill F, eds. Rapid Cycle Real-Time PCR Methods and Applications. Springer-Verlag, 2002:3-27; Zbinden R: Aggregatibacter, Capnocytophaga, Eikenella, Kingella, Pasteurella, and other fastidious or rarely encountered gram-negative rods. In: Carroll K, Pfaller M, eds. Manual of Clinical Microbiology. 12th ed. ASM Press; 2019:656-669)
Day(s) Performed
Monday through Friday
Report Available
2 to 7 daysSpecimen Retention Time
1 weekPerforming Laboratory
Mayo Clinic Laboratories in Rochester
Test Classification
This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. It has not been cleared or approved by the US Food and Drug Administration.CPT Code Information
87798
LOINC Code Information
| Test ID | Test Order Name | Order LOINC Value |
|---|---|---|
| KKRP | Kingella kingae PCR | 65809-6 |
| Result ID | Test Result Name | Result LOINC Value |
|---|---|---|
| KKSRC | Specimen Source | 31208-2 |
| 48324 | Kingella kingae PCR | 65809-6 |
Forms
If not ordering electronically, complete, print, and send a Microbiology Test Request (T244)with the specimen.