Department of Respiratory Medicine, Sydney Children's Hospital, Randwick, Australia.
Respirology. 2012 Jan;17(1):114-9. doi: 10.1111/j.1440-1843.2011.02035.x.
National surveillance of invasive pneumococcal disease (IPD) includes serotyping Streptococcus pneumoniae (SP) isolates from sterile site cultures. PCR is more sensitive and can identify more SP serotypes (STs) in culture-negative samples. The aim of this study was to determine whether enhanced surveillance of childhood empyema, using PCR, provides additional serotype information compared with conventional surveillance.
Pleural fluid (PF) from children with empyema were cultured and tested by PCR to identify SP, targeting the autolysin gene (lytA). Multiplex PCR-based reverse line blot assay was used to identify SP STs. Corresponding IPD surveillance and serotype data were obtained from the National Notifiable Diseases Surveillance System (NNDSS).
Eighty-nine children with empyema, aged ≤16 years, were recruited between April 2008 and March 2009, inclusive. SP was isolated from 5/84 (5.9%) PF cultures and by PCR in 43/79 (54.4%) PF samples. Serotypes were unidentifiable in 15 samples. The frequency of six serotypes (or serotype pairs) identified in 28 samples, including one with two serotypes, were: ST1, n = 4/29 (13.8%); ST3, n = 9/29 (31.0%); ST19A, n = 12/29 (41.4%); ST7F/7A, n = 1/29 (3.4%); ST9V/9A, n = 1/29 (3.4%); ST22F/22A, n = 2/29 (6.9%). Over the same period, 361 IPD patients, aged 16 years or less, were notified to NNDSS. Among 331 serotypeable NNDSS isolates (71.5% from blood), the frequencies of ST1 and 3 were significantly lower than in PF samples: ST1, n = 8/331 (2.4%; P < 0.05); ST3, n = 13/331 (3.9%; P < 0.0001).
The use of PCR to identify and serotype SP in culture-negative specimens provides additive information.
全国侵袭性肺炎球菌病(IPD)监测包括对无菌部位培养物中肺炎链球菌(SP)分离株进行血清型分型。PCR 更敏感,可在培养阴性样本中鉴定出更多 SP 血清型(ST)。本研究旨在确定与传统监测相比,使用 PCR 增强对儿童脓胸的监测是否提供了额外的血清型信息。
收集年龄≤16 岁的脓胸患儿的胸腔积液(PF)并进行培养和 PCR 检测,以鉴定 SP,检测靶点为自溶素基因(lytA)。采用基于多重 PCR 的反向线印迹检测法鉴定 SP ST。从国家法定传染病监测系统(NNDSS)中获得相应的 IPD 监测和血清型数据。
2008 年 4 月至 2009 年 3 月期间,共纳入 89 例年龄≤16 岁的脓胸患儿。5/84(5.9%)份 PF 培养物和 43/79(54.4%)份 PF 样本中通过 PCR 分离出 SP。15 份样本无法确定血清型。28 份样本中鉴定出 6 种血清型(或血清型对)的频率,包括两种血清型各 1 份:ST1,n=4/29(13.8%);ST3,n=9/29(31.0%);ST19A,n=12/29(41.4%);ST7F/7A,n=1/29(3.4%);ST9V/9A,n=1/29(3.4%);ST22F/22A,n=2/29(6.9%)。同期,NNDSS 报告了 361 例 16 岁及以下的 IPD 患者。在 331 份可分型的 NNDSS 分离株(71.5%来自血液)中,ST1 和 ST3 的频率明显低于 PF 样本:ST1,n=8/331(2.4%;P<0.05);ST3,n=13/331(3.9%;P<0.0001)。
使用 PCR 鉴定和血清型分型培养阴性标本可提供附加信息。