Download PDF
J Pediatr Infect Dis 2018; 13(03): 216-223
DOI: 10.1055/s-0038-1641603
Original Article
Georg Thieme Verlag KG Stuttgart · New York

Real-Time PCR Assay for Detection of Kingella kingae in Children

Theresa Madigan
1   Division of Pediatric Infectious Diseases, Department of Pediatric and Adolescent Medicine, Mayo Clinic, Rochester, Minnesota, United States
,
Scott A. Cunningham
2   Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, United States
,
Poornima Ramanan
2   Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, United States
,
Micah M. Bhatti
3   Clinical Microbiology Laboratory, Department of Laboratory Medicine, The University of Texas, MD Anderson Cancer Center, Houston, Texas, United States
,
Robin Patel
2   Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, United States
4   Division of Infectious Diseases, Department of Medicine, Mayo Clinic, Rochester, Minnesota, United States
› Author Affiliations Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
Further Information

Publication History

03 January 2018

02 March 2018

Publication Date:
11 April 2018 (online)

Also available at
    Permissions and Reprints

Abstract

Background Kingella kingae is a known cause of osteoarticular infections in children younger than 4 years of age, but it is not always recoverable in culture. Molecular methods are increasingly used for diagnosis.

Methods To facilitate diagnosis of K. kingae septic arthritis, we developed a real-time polymerase chain reaction (PCR) assay for the detection of K. kingae that targets the repeat-in-toxin gene (rtxB).

Results We present three pediatric patients with K. kingae septic arthritis at our institution who were diagnosed using the real-time PCR assay. All underwent arthrotomy with irrigation and debridement and were symptom-free after 3 weeks of therapy with β-lactam antibiotics. Cultures of synovial fluid or tissue grew K. kingae in two of three; K. kingae real-time PCR was positive in all three patients. In addition, 11 cases of K. kingae osteoarticular infection were diagnosed through Mayo Medical Laboratories using this assay. The limit of detection of the real-time PCR assay was 73.7 colony-forming unit (CFU)/µL for tissue and 1.3 CFU/µL for synovial fluid.

Conclusions PCR-based detection methods are faster and more sensitive than conventional culture-based methods for the diagnosis of K. kingae osteoarticular infections in children.

Keywords

Kingella kingae - real-time PCR - children

Note

This research was presented in part at the St. Jude Pediartic Infectious Diseases (PIDS) Research Conference held at Memphis, TN, March 2018.


 

  • References

  • 1 Yagupsky P. Kingella kingae: from medical rarity to an emerging paediatric pathogen. Lancet Infect Dis 2004; 4 (06) 358-367
    CrossrefPubMedSearch in Google Scholar
  • 2 Henriksen SD, Bovre K. Moraxella kingii sp. nov., a haemolytic, saccharolytic species of the genus Moraxella . J Gen Microbiol 1968; 51 (03) 377-385
    CrossrefPubMedSearch in Google Scholar
  • 3 Yagupsky P, Porsch E, St Geme III JW. Kingella kingae: an emerging pathogen in young children. Pediatrics 2011; 127 (03) 557-565
    CrossrefPubMedSearch in Google Scholar
  • 4 Saphyakhajon P, Greene G. Kingella kingae: an emerging pathogen of acute osteoarticular infections in children. Pediatrics 2006; 117 (01) 249 , author reply 249–250
    PubMedSearch in Google Scholar
  • 5 Yagupsky P, Dagan R. Kingella kingae: an emerging cause of invasive infections in young children. Clin Infect Dis 1997; 24 (05) 860-866
    CrossrefPubMedSearch in Google Scholar
  • 6 Lundy DW, Kehl DK. Increasing prevalence of Kingella kingae in osteoarticular infections in young children. J Pediatr Orthop 1998; 18 (02) 262-267
    PubMedSearch in Google Scholar
  • 7 Ilharreborde B, Bidet P, Lorrot M. , et al. New real-time PCR-based method for Kingella kingae DNA detection: application to samples collected from 89 children with acute arthritis. J Clin Microbiol 2009; 47 (06) 1837-1841
    CrossrefPubMedSearch in Google Scholar
  • 8 Chometon S, Benito Y, Chaker M. , et al. Specific real-time polymerase chain reaction places Kingella kingae as the most common cause of osteoarticular infections in young children. Pediatr Infect Dis J 2007; 26 (05) 377-381
    CrossrefPubMedSearch in Google Scholar
  • 9 Ceroni D, Cherkaoui A, Ferey S, Kaelin A, Schrenzel J. Kingella kingae osteoarticular infections in young children: clinical features and contribution of a new specific real-time PCR assay to the diagnosis. J Pediatr Orthop 2010; 30 (03) 301-304
    CrossrefPubMedSearch in Google Scholar
  • 10 Ferroni A, Al Khoury H, Dana C. , et al. Prospective survey of acute osteoarticular infections in a French paediatric orthopedic surgery unit. Clin Microbiol Infect 2013; 19 (09) 822-828
    CrossrefPubMedSearch in Google Scholar
  • 11 Yagupsky P, Dagan R, Howard CW, Einhorn M, Kassis I, Simu A. High prevalence of Kingella kingae in joint fluid from children with septic arthritis revealed by the BACTEC blood culture system. J Clin Microbiol 1992; 30 (05) 1278-1281
    CrossrefPubMedSearch in Google Scholar
  • 12 Al-Qwbani M, Jiang N, Yu B. Kingella kingae-associated pediatric osteoarticular infections: an overview of 566 reported cases. Clin Pediatr (Phila) 2016; 55 (14) 1328-1337
    CrossrefPubMedSearch in Google Scholar
  • 13 Slinger R, Moldovan I, Bowes J, Chan F. Polymerase chain reaction detection of Kingella kingae in children with culture-negative septic arthritis in eastern Ontario. Paediatr Child Health 2016; 21 (02) 79-82
    CrossrefPubMedSearch in Google Scholar
  • 14 Cherkaoui A, Ceroni D, Emonet S, Lefevre Y, Schrenzel J. Molecular diagnosis of Kingella kingae osteoarticular infections by specific real-time PCR assay. J Med Microbiol 2009; 58 (Pt 1): 65-68
    CrossrefPubMedSearch in Google Scholar
  • 15 Verdier I, Gayet-Ageron A, Ploton C. , et al. Contribution of a broad range polymerase chain reaction to the diagnosis of osteoarticular infections caused by Kingella kingae: description of twenty-four recent pediatric diagnoses. Pediatr Infect Dis J 2005; 24 (08) 692-696
    CrossrefPubMedSearch in Google Scholar
  • 16 Rosey AL, Abachin E, Quesnes G. , et al. Development of a broad-range 16S rDNA real-time PCR for the diagnosis of septic arthritis in children. J Microbiol Methods 2007; 68 (01) 88-93
    CrossrefPubMedSearch in Google Scholar
  • 17 Arnold JC, Bradley JS. Osteoarticular infections in children. Infect Dis Clin North Am 2015; 29 (03) 557-574
    CrossrefPubMedSearch in Google Scholar
  • 18 Moumile K, Merckx J, Glorion C, Berche P, Ferroni A. Osteoarticular infections caused by Kingella kingae in children: contribution of polymerase chain reaction to the microbiologic diagnosis. Pediatr Infect Dis J 2003; 22 (09) 837-839
    PubMedSearch in Google Scholar
  • 19 Levy PY, Fournier PE, Fenollar F, Raoult D. Systematic PCR detection in culture-negative osteoarticular infections. Am J Med 2013; 126 (12) 1143.e25-1143.e33
    CrossrefPubMedSearch in Google Scholar
  • 20 Haldar M, Butler M, Quinn CD, Stratton CW, Tang YW, Burnham CA. Evaluation of a real-time PCR assay for simultaneous detection of Kingella kingae and Staphylococcus aureus from synovial fluid in suspected septic arthritis. Ann Lab Med 2014; 34 (04) 313-316
    CrossrefPubMedSearch in Google Scholar
  • 21 Lehours P, Freydière AM, Richer O. , et al. The rtxA toxin gene of Kingella kingae: a pertinent target for molecular diagnosis of osteoarticular infections. J Clin Microbiol 2011; 49 (04) 1245-1250
    CrossrefPubMedSearch in Google Scholar
  • 22 Yang S, Rothman RE. PCR-based diagnostics for infectious diseases: uses, limitations, and future applications in acute-care settings. Lancet Infect Dis 2004; 4 (06) 337-348
    CrossrefPubMedSearch in Google Scholar
  • 23 Binnicker MJ, Buckwalter SP, Eisberner JJ. , et al. Detection of Coccidioides species in clinical specimens by real-time PCR. J Clin Microbiol 2007; 45 (01) 173-178
    CrossrefPubMedSearch in Google Scholar
  • 24 El Houmami N, Bakour S, Bzdrenga J. , et al. Isolation and characterization of Kingella negevensis sp. nov., a novel Kingella species detected in a healthy paediatric population. Int J Syst Evol Microbiol 2017; 67 (07) 2370-2376
    CrossrefPubMedSearch in Google Scholar
  • 25 Kehl-Fie TE, St Geme III JW. Identification and characterization of an RTX toxin in the emerging pathogen Kingella kingae . J Bacteriol 2007; 189 (02) 430-436
    CrossrefPubMedSearch in Google Scholar
  • 26 Luhmann JD, Luhmann SJ. Etiology of septic arthritis in children: an update for the 1990s. Pediatr Emerg Care 1999; 15 (01) 40-42
    CrossrefPubMedSearch in Google Scholar
  • 27 Lyon RM, Evanich JD. Culture-negative septic arthritis in children. J Pediatr Orthop 1999; 19 (05) 655-659
    PubMedSearch in Google Scholar
  • 28 Saphyakhajon P, Joshi AY, Huskins WC, Henry NK, Boyce TG. Empiric antibiotic therapy for acute osteoarticular infections with suspected methicillin-resistant Staphylococcus aureus or Kingella . Pediatr Infect Dis J 2008; 27 (08) 765-767
    CrossrefPubMedSearch in Google Scholar
  • 29 Yagupsky P. Antibiotic susceptibility of Kingella kingae isolates from children with skeletal system infections. Pediatr Infect Dis J 2012; 31 (02) 212
    PubMedSearch in Google Scholar
  • 30 Yagupsky P, Katz O, Peled N. Antibiotic susceptibility of Kingella kingae isolates from respiratory carriers and patients with invasive infections. J Antimicrob Chemother 2001; 47 (02) 191-193
    CrossrefPubMedSearch in Google Scholar
  • 31 Sordillo EM, Rendel M, Sood R, Belinfanti J, Murray O, Brook D. Septicemia due to beta-lactamase-positive Kingella kingae . Clin Infect Dis 1993; 17 (04) 818-819
    CrossrefPubMedSearch in Google Scholar
  • 32 Yagupsky P, Slonim A, Amit U, Porat N, Dagan R. Beta-lactamase production by Kingella kingae in Israel is clonal and common in carriage organisms but rare among invasive strains. Eur J Clin Microbiol Infect Dis 2013; 32 (08) 1049-1053
    CrossrefPubMedSearch in Google Scholar
  • 33 Basmaci R, Bidet P, Berçot B. , et al. First identification of a chromosomally located penicillinase gene in Kingella kingae species isolated in continental Europe. Antimicrob Agents Chemother 2014; 58 (10) 6258-6259
    CrossrefPubMedSearch in Google Scholar
  • 34 Basmaci R, Bonacorsi S, Bidet P. , et al. Genotyping, local prevalence and international dissemination of β-lactamase-producing Kingella kingae strains. Clin Microbiol Infect 2014; 20 (11) O811-O817
    CrossrefPubMedSearch in Google Scholar
  • 35 Banerjee A, Kaplan JB, Soherwardy A. , et al. Characterization of TEM-1 beta-lactamase-producing Kingella kingae clinical isolates. Antimicrob Agents Chemother 2013; 57 (09) 4300-4306
    CrossrefPubMedSearch in Google Scholar
  • 36 Birgisson H, Steingrimsson O, Gudnason T. Kingella kingae infections in paediatric patients: 5 cases of septic arthritis, osteomyelitis and bacteraemia. Scand J Infect Dis 1997; 29 (05) 495-498
    CrossrefPubMedSearch in Google Scholar
  • 37 Yagupsky P, Merires M, Bahar J, Dagan R. Evaluation of novel vancomycin-containing medium for primary isolation of Kingella kingae from upper respiratory tract specimens. J Clin Microbiol 1995; 33 (05) 1426-1427
    CrossrefPubMedSearch in Google Scholar
  • 38 El Houmami N, Schrenzel J, Yagupsky P. , et al. Draft genome sequence of Kingella negevensis SW7208426, the first European strain of K. negevensis isolated from a healthy child in Switzerland. Genome Announc 2017; 5 (26) e00571-17
    CrossrefPubMedSearch in Google Scholar
  • 39 El Houmami N, Bzdrenga 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; 55 (10) 3113-3122
    CrossrefPubMedSearch in Google Scholar
  • 40 Opota O, Laurent S, Pillonel T. , et al. Genomics of the new species Kingella negevensis: diagnostic issues and identification of a locus encoding a RTX toxin. Microbes Infect 2017; 19 (11) 546-552
    CrossrefPubMedSearch in Google Scholar