양성혈액배양에서 메치실린 내성 황색 포도알균의 직접검출을 위한 PBP2a 라텍스 응집법, PBP2a 신속 면역크로마토그래픽법 및 Chromogenic 배지법의 비교
Direct Detection of Methicillin-Resistant Staphylococcus aureus from Blood Cultures Using Three Non-Molecular Methods: PBP2a Latex Agglutination, PBP2a Rapid Immunochromatographic Assay and MRSA-Chromogenic Medium
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배경: Staphylococcus aureus를 포함하는 혈액배양에서 직접 methicillin 내성의 검출을 위한 3가지 표현학적 방법인 penicillin-binding protein 2a (PBP2a) 라텍스 응집법, PBP2a immunochromatographic assay (ICA) 및 MRSA chromogenic medium (MRSA-CM)의 성능을 비교하였다. 방법: nuc PCR과 mecA PCR로 확인된 50주의 methicillin-resistant S. aureus (MRSA)와 50주의 methicillin-susceptible S. aureus (MSSA)를 혈액배양병에 넣고 균의 증식 신호가 나타나면 5 mL의 혈액을 취하여 혈청분리 튜브에 옮긴 후 1,300g에서 10분간 원심분리하여 균 침사를 얻었다. 균 침사를 PBP2a 라텍스 응집법, MRSA-CM, 및 PBP2a ICA 검사에 이용하였다. 추가로 혈액한천 배지에서 자란 균집락을 이용하여 PBP2a LA 검사를 시행하였다. 결과: 혈액배양병에서 직접 MRSA의 검출을 위한 EZ-Step MRSA rapid kit와 MRSA-CM의 예민도와 특이도는 각각 98%와 100% 그리고 100%와 100%이었다. 집락을 이용한 PBP2a LA 검사는 mecA PCR과 완전히 일치하였으나 혈액배양병에서 얻은 침사를 직접 이용한 PBP2a 라텍스 응집법에서 7주의 MRSA와 5주의 MSSA는 불분명한 응집을 보였으며, 불분명한 응집을 음성과 양성으로 각각 판단하였을 때의 예민도와 특이도는 각각 78%와 100% 그리고 90%와 92%이었다. 결론: 혈액배양에서 MRSA를 직접 검출하는 데 있어, EZ-Step MRSA rapid kit와 MRSA-CM은 매우 우수한 결과를 보였다. 제조사의 지침대로 균집락을 이용한 PBP2a 라텍스 응집법은 MRSA를 감별하는 데 이전 두 검사방법과 같이 매우 우수한 결과를 보였으나, 혈액배양에서 직접 검출할 때는 일부에서 불분명한 응집을 보였다.
Abstract
Background: This study compared three non-molecular methods for the detection of methicillin-resistance directly from blood cultures containing Staphylococcus aureus: penicillin-binding protein (PBP) 2a latex agglutination (LA), PBP2a immunochromatographic assay (ICA) and MRSA chromogenic medium (CM). Methods: Fifty methicillin-resistant S. aureus (MRSA) and 50 methicillin-susceptible S. aureus (MSSA) were seeded into blood-culture bottles. When isolates returned a positive signal, 5 mL of culture was added to serum separator tubes and centrifuged at 1,300 g for 10 min. The pellets were then used as the inoculum for the PBP2a LA, MRSA-CM and PBP2a ICA. The pure colony was used for PBP2a LA test, additionally. Results: The respective sensitivities and specificities were 98 and 100% for PBP2a ICA, and 100 and 100% for MRSA-CM in direct detection of MRSA from positive blood culture. The results of PBP2a LA test using pure colony were entirely compatible with those by mecA gene PCR but the PBP2a LA test using the pellets directly isolated from positive blood culture showed sometimes ambiguous agglutination; its sensitivity and specificity were 78 and 100%, if ambiguous results were scored as negative, and were 90 and 92%, if ambiguous results were scored as positive, respectively. Conclusion: For direct detection of MRSA in positive blood culture, MRSA-CM and PBP2a ICA provided excellent results. The PBP2a LA test using pure colony also gave excellent results but the PBP2a LA test by the direct method using pellet of positive blood culture was slightly less sensitive than the other two methods.
참고문헌 (20)
- Lodise TP, "Clinical and economic impact of methicillin resistance in patients with Staphylococcus aureus bacteremia", Diagn Microbiol Infect Dis, 52, pp.113~122 (2005)
- Colakoglu S, "Performance of MRSA ID chromogenic medium for detection of methicillin-resistant Staphylococcus aureus directly from blood cultures and clinical specimens", Diagn Microbiol Infect Dis, 59, pp.319~323 (2007)
- Pape J, "Use of BBL CHROMagar MRSA medium for identification of methicillin-resistant Staphylococcus aureus directly from blood cultures", J Clin Microbiol, 44, pp.2575~2576 (2006)
- Harriau P, "Use of BioRad plating agar MRSASelect for the daily detection of methicillin resistant staphylococci isolated from samples taken from blood culture bottles", Pathol Biol (Paris), 54, pp.506~509 (2006)
- Bennett K, "Rapid differentiation of methicillin-resistant Staphylococcus aureus and methicillin-susceptible Staphylococcus aureus from blood cultures by use of a direct cefoxitin disk diffusion test", J Clin Microbiol, 46, pp.3836~3838 (2008)
- Chapin KC, "Evaluation of penicillin binding protein 2a latex agglutination assay for identification of methicillin- resistant Staphylococcus aureus directly from blood cultures", J Clin Microbiol, 42, pp.1283~1284 (2004)
- Qian Q, "Direct detection of methicillin resistance in Staphylococcus aureus in blood culture broth by use of a penicillin binding protein 2a latex agglutination test", J Clin Microbiol, 48, pp.1420~1421 (2010)
- Louie L, "Rapid detection of methicillin-resistant staphylococci from blood culture bottles by using a multiplex PCR assay", J Clin Microbiol, 40, pp.2786~2790 (2002)
- Shrestha NK, "Rapid identification of Staphylococcus aureus and the mecA gene from BacT", ALERT blood culture bottles by using the LightCycler system, 2002, pp.40, 2659~2661
- Wolk DM, "Rapid detection of Staphylococcus aureus and methicillinresistant S. aureus (MRSA) in wound specimens and blood cultures: multicenter preclinical evaluation of the Cepheid Xpert MRSA", SA skin and soft tissue and blood culture assays, 2009, pp.47, 823~826
- Shin KS, "Direct detection of methicillin-resistant Staphylococcus aureus from blood cultures using an immunochromatographic immunoassaybased MRSA rapid kit for the detection of penicillin-binding protein 2a", Diagn Microbiol Infect Dis, 67, pp.301~303 (2010)
- Brakstad OG, "Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene", J Clin Microbiol, 30, pp.1654~1660 (1992)
- Arbique J, "Comparison of the Velogene Rapid MRSA Identification Assay, Denka MRSA-Screen Assay, and BBL Crystal MRSA ID System for rapid identification of methicillin-resistant Staphylococcus aureus", Diagn Microbiol Infect Dis, 40, pp.5~10 (2001)
- Yamazumi T, "Jones RN. Comparison of the Vitek Gram-Positive Susceptibility 106 card and the MRSA-screen latex agglutination test for determining oxacillin resistance in clinical bloodstream isolates of Staphylococcus aureus", J Clin Microbiol, 39, pp.53~56 (2001)
- Bassiwa L, "Direct detection of altered penicillin binding protein (PBP2’) in Staphylococcus aureus and coagulasenegative staphylococci in blood cultures", Am. Soc. Microbiol. (2003)
- Shovlin A, "Direct detection of methicillin resistance Staphylococcus aureus in blood culture bottles", Am. Soc. Microbiol (2004)
- Yamazumi T, "Rapid detection of methicillin resistance in clinical isolates of Staphylococcus spp. directly from blood cultures using a PBP2’ slide agglutination test", Am Soc. Microbiol (2002)
- Diederen B, "Performance of CHROMagar MRSA medium for detection of methicillin-resistant Staphylococcus aureus", J Clin Microbiol, 43, pp.1925~1927 (2005)
- Hedin G, "Evaluation of two new chromogenic media, CHROMagar MRSA and S. aureus ID, for identifying Staphylococcus aureus and screening methicillin-resistant S. aureus", J Clin Microbiol, 43, pp.4242~4244 (2005)
- Stoakes L, "Prospective comparison of a new chromogenic medium, MRSASelect, to CHROMagar MRSA and mannitol-salt medium supplemented with oxacillin or cefoxitin for detection of methicillin-resistant Staphylococcus aureus", J Clin Microbiol, 44, pp.637~639 (2006)