评估双靶标实时 PCR 检测丹麦儿科人群中金氏金菌的应用。

Evaluation of dual target-specific real-time PCR for the detection of Kingella kingae in a Danish paediatric population.

机构信息

a Department of Clinical Microbiology , Copenhagen University Hospital Hvidovre , Hvidovre , Denmark.

b Department of Clinical Medicine , University of Copenhagen , Copenhagen , Denmark.

出版信息

Infect Dis (Lond). 2018 Mar;50(3):200-206. doi: 10.1080/23744235.2017.1376254. Epub 2017 Sep 15.

DOI:10.1080/23744235.2017.1376254

PMID:28914110

Abstract

BACKGROUND

We aimed to evaluate the relevance of dual target real-time polymerase chain (PCR) assays targeting the rtxA and cpn60 genes of the paediatric pathogen Kingella kingae. We also studied for the first time the clinical and epidemiological features of K. kingae infections in a Danish population.

METHOD

Children with K. kingae-positive cultures were identified from 11,477 children and 86 children younger than 16 years old from whom blood cultures and joint fluid cultures were obtained between January 2010 and November 2016. Results were then compared to microbiological results obtained from 29 joint fluids (28 children) tested by dual target K. kingae real-time PCR from September 2014 to November 2016. Epidemiological data of all children with microbiologically confirmed K. kingae infections were collected.

RESULTS

From 2010 to 2016, we diagnosed 17 children with microbiological-proven K. kingae infections. During this period, blood cultures from five children and joint fluid cultures from a single child yielded K. kingae. Dual target K. kingae real-time PCR allowed us to increase the diagnostic yield of K. kingae infections by detecting the organism in 12 of 29 (41.4%) specimens. Notably, the 12 real-time PCR-positive specimens were rtxA-positive whereas only 10 (83.3%) were cpn60-positive. PCR-positive children were significantly younger than PCR-negative children (p-value: .01). A significant seasonal variation was found for patients with proven K. kingae infection (p-value: <.001), with a peak in autumn.

CONCLUSION

Dual target-specific real-time PCR markedly improved the detection of K. kingae in clinical specimens when compared to culture methods.

摘要

背景

我们旨在评估针对小儿病原体金氏金氏菌 rtxA 和 cpn60 基因的双靶实时聚合酶链（PCR）检测的相关性。我们还首次研究了丹麦人群中金氏金氏菌感染的临床和流行病学特征。

方法

从 2010 年 1 月至 2016 年 11 月，从 11477 名儿童和 86 名 16 岁以下儿童中鉴定出金氏金氏菌培养阳性的儿童，从这些儿童中获得了血培养和关节液培养。然后将结果与 2014 年 9 月至 2016 年 11 月期间通过双靶金氏金氏菌实时 PCR 检测的 29 份关节液（28 名儿童）的微生物学结果进行比较。收集了所有经微生物学证实的金氏金氏菌感染儿童的流行病学数据。

结果

从 2010 年到 2016 年，我们诊断出 17 名儿童患有微生物学证实的金氏金氏菌感染。在此期间，5 名儿童的血培养和 1 名儿童的关节液培养中均培养出金氏金氏菌。双靶金氏金氏菌实时 PCR 检测可将金氏金氏菌感染的诊断率提高 12/29（41.4%）。值得注意的是，12 份实时 PCR 阳性标本 rtxA 阳性，而仅 10 份（83.3%） cpn60 阳性。PCR 阳性儿童明显比 PCR 阴性儿童年龄小（p 值：<.01）。发现证实有金氏金氏菌感染的患者有明显的季节性变化（p 值：<.001），秋季达到高峰。