Pediatric Clinic 1, Department of Pathophysiology and Transplantation, Università degli Studi di Milano, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Via Commenda 9, 20122 Milano, Italy.
Eur J Clin Microbiol Infect Dis. 2013 Sep;32(9):1129-33. doi: 10.1007/s10096-013-1856-2. Epub 2013 Mar 17.
The purpose of this investigation was to evaluate the effectiveness of posterior pharyngeal and nasopharyngeal swabs in identifying and quantifying meningococcal carriage. Two swab samples were obtained from 564 healthy adolescents aged 15-19 years, the first taken from the posterior pharyngeal wall through the mouth and the second through the nose. Bacterial genomic DNA was extracted and screened for Neisseria meningitidis by means of two separate singleplex real-time polymerase chain reactions (real-time PCRs) in order to identify the CtrA and sodC genes. Subsequently, N. meningitidis-positive samples underwent a further singleplex real-time PCR in order to determine the N. meningitidis serogroup, and the DNA was quantified by means of standard curves. Thirty-seven subjects (6.6 %) were found to be carriers of N. meningitidis. The most frequently carried serogroup was serogroup B (15 cases, 40.5 %); serogroups A, Y, X, W135 and Z were found in, respectively, two (5.4 %), five (13.5 %), four (10.8 %), three (8.1 %) and one subject (2.7 %); the serogroup was not identified in seven cases. The detection of carrier status was significantly more frequent using posterior pharyngeal swabs (5.3 % vs. 2.1 %; p = 0.004), which also contained a significantly larger number of N. meningitidis genomic copies (4.91 ± 1.39 vs. 2.50 ± 0.8 log10 genomic copies/mL; p < 0.001). Posterior pharyngeal swabs seem to be better than nasopharyngeal swabs for detecting N. meningitidis carriage in large-scale epidemiological studies because they identify a significantly larger number of pathogen carriers and recover a significantly larger amount of bacterial DNA.
本研究旨在评估咽后和鼻咽拭子在鉴定和定量脑膜炎奈瑟菌带菌中的有效性。从 564 名 15-19 岁的健康青少年中采集了两份拭子样本,第一份通过口腔从咽后壁采集,第二份通过鼻腔采集。提取细菌基因组 DNA,并通过两种单独的实时聚合酶链反应(real-time PCR)筛选脑膜炎奈瑟菌,以鉴定 CtrA 和 sodC 基因。随后,对脑膜炎奈瑟菌阳性样本进行进一步的单重实时 PCR,以确定脑膜炎奈瑟菌血清群,并通过标准曲线定量 DNA。发现 37 名受试者(6.6%)携带脑膜炎奈瑟菌。携带最频繁的血清群是 B 群(15 例,40.5%);血清群 A、Y、X、W135 和 Z 分别在 2 例(5.4%)、5 例(13.5%)、4 例(10.8%)、3 例(8.1%)和 1 例(2.7%)中发现;7 例无法确定血清群。咽后拭子检测带菌状态的频率明显更高(5.3% vs. 2.1%;p=0.004),其中脑膜炎奈瑟菌基因组拷贝数也明显更多(4.91±1.39 对 2.50±0.8 log10 基因组拷贝/mL;p<0.001)。在大规模的流行病学研究中,咽后拭子似乎比鼻咽拭子更能检测到脑膜炎奈瑟菌的带菌状态,因为它们能识别出更多的病原菌携带者,并能回收更多的细菌 DNA。