Chometon Sylvia, Benito Yvonne, Chaker Mourad, Boisset Sandrine, Ploton Christine, Bérard Jérôme, Vandenesch François, Freydiere Anne Marie
Laboratoire de Bactériologie, Hôpital Cardiologique Louis Pradel, Lyon, France.
Pediatr Infect Dis J. 2007 May;26(5):377-81. doi: 10.1097/01.inf.0000259954.88139.f4.
The use of universal 16S rDNA polymerase chain reaction (PCR) has recently shown that the place of Kingella kingae in osteoarticular infections (OAI) in young children has been underestimated, but this technique is not the most sensitive or the most rapid method for molecular diagnosis. We developed a specific real-time PCR method to detect K. kingae DNA and applied it to the etiologic diagnosis of OAI.
All children admitted to a pediatric unit for OAI between January 2004 and December 2005 were enrolled in this prospective study. Culture-negative osteoarticular specimens were tested by 16S rDNA PCR and by K. kingae-specific real-time PCR when sufficient sample remained.
By culture alone, a pathogen was identified in 45% of the 131 specimens tested (Staphylococcus aureus, n = 25; K. kingae, n = 17; others, n = 18). 16S rDNA PCR and K. kingae-specific PCR were both applied to 61 of the culture-negative samples. The combination of culture and 16S rDNA PCR identified a pathogen in 61% of cases (K. kingae DNA, n = 16; DNA of other microorganisms, n = 5). Specific real-time PCR identified a further 6 cases caused by K. kingae and confirmed all 16 universal PCR-positive cases, bringing the overall documentation rate to 66%. K. kingae was the leading cause of OAI in this pediatric series (n = 39, 45%), followed by S. aureus (n = 25, 29%)
The K. kingae-specific real-time PCR places K. kingae as the leading cause of OAI in children at our hospital.
最近采用通用的16S核糖体DNA聚合酶链反应（PCR）显示，金氏金杆菌在幼儿骨关节炎感染（OAI）中的地位一直被低估，但该技术并非分子诊断中最敏感或最快速的方法。我们开发了一种特异性实时PCR方法来检测金氏金杆菌DNA，并将其应用于OAI的病因诊断。
2004年1月至2005年12月期间因OAI入住儿科病房的所有儿童均纳入本前瞻性研究。当有足够样本时，对培养阴性的骨关节炎标本进行16S rDNA PCR和金氏金杆菌特异性实时PCR检测。
仅通过培养，在131份检测标本中有45%鉴定出病原体（金黄色葡萄球菌，n = 25；金氏金杆菌，n = 17；其他，n = 18）。16S rDNA PCR和金氏金杆菌特异性PCR均应用于61份培养阴性的样本。培养与16S rDNA PCR相结合在61%的病例中鉴定出病原体（金氏金杆菌DNA，n = 16；其他微生物DNA，n = 5）。特异性实时PCR又鉴定出6例由金氏金杆菌引起的病例，并确认了所有16例通用PCR阳性病例，使总体确诊率达到66%。在这个儿科系列中，金氏金杆菌是OAI的主要病因（n = 39，45%），其次是金黄色葡萄球菌（n =
