Alternative Molecular Methods for Improved Detection of Meningococcal Carriage and Measurement of Bacterial Density
Alternative Molecular Methods for Improved Detection of Meningococcal Carriage and Measurement of Bacterial Density
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DOI：
10.1128/jcm.01428-16
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发表时间：
2016-11-01
影响因子：
9.4
通讯作者：
Greenwood, Brian M.
中科院分区：
文献类型：
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作者：
Manigart, Olivier;Okeakpu, Jacinta;Greenwood, Brian M.
检测脑膜炎奈瑟菌咽部携带的常规方法是复杂的。需要具有改进的性能的更简单的方法。我们研究了两种替代方法。从冈比亚999名10至18岁的学生中收集了三份咽拭子。运输N。使用三种不同的方法研究脑膜炎：（i）在Thayer-Martin选择性培养基上铺板，并通过常规微生物方法进行检测，然后进行PCR检测;（ii）接种在Todd-Hewitt肉汤（THB）中，培养过夜后，通过PCR检测;和（iii）将拭子压缩在滤纸上，并在DNA浓缩后，通过PCR检测。在THB培养后进行PCR检测，其灵敏度是常规方法的2倍以上。脑膜炎（13.2%比5.7%; P < 0.0001）。从滤纸上提取DNA后进行PCR的敏感性与常规方法相似（4.9%对5.7%; P = 0.33）。经肉汤培养法检测，有21株脑膜炎球菌为W基因组，12株为B基因组，8株为Y基因组，3株为E基因组，2株为X基因组，68株脑膜炎球菌具有无荚膜基因间区。三种方法的基因组和无囊生物的分布相似。从滤纸中提取的样品中的携带密度范围为1至25，000个DNA拷贝。过夜生长的肉汤培养物的PCR使N.脑膜炎病毒携带分离株与常规方法的比较。这种方法可以提高运输研究的效率。在滤纸上收集，然后进行定量PCR，可用于在资源有限的地区进行密度测量和携带研究。
Conventional methods for detecting pharyngeal carriage of Neisseria meningitidis are complex. There is a need for simpler methods with improved performance. We have investigated two alternative approaches. Three pharyngeal swabs were collected from 999 pupils aged 10 to 18 years in The Gambia. Carriage of N. meningitidis was investigated by using three different methods: (i) plating on Thayer-Martin selective medium and testing by conventional microbiological methods followed by PCR testing; (ii) seeding in Todd-Hewitt broth (THB) and, after culture overnight, testing by PCR; and (iii) compression of the swab on filter paper and, after DNA concentration, testing by PCR. PCR after culture in THB was more than twice as sensitive as conventional methods in detecting N. meningitidis (13.2% versus 5.7%; P < 0.0001). PCR after DNA extraction from filter paper had a sensitivity similar to that of conventional methods (4.9% versus 5.7%; P = 0.33). Capsular genogroups detected by broth culture were genogroups W(21 isolates), B (12 isolates), Y (8 isolates), E (3 isolates), and X (2 isolates), and 68 meningococci had the capsule-null intergenic region. The distributions of genogroups and of capsule-null organisms were similar with each of the three methods. The carriage density in samples extracted from filter paper ranged from 1 to 25,000 DNA copies. PCR of broth cultures grown overnight doubled the yield of N. meningitidis carriage isolates compared with conventional methods. This approach could improve the efficiency of carriage studies. Collection on filter paper followed by quantitative PCR could be useful for density measurement and for carriage studies in areas with limited resources.