高通量基于信息单核苷酸多态性的淋病奈瑟菌测序酶联质谱 iPLEX 平台分型。
High-throughput informative single nucleotide polymorphism-based typing of Neisseria gonorrhoeae using the Sequenom MassARRAY iPLEX platform.
机构信息
Queensland Paediatric Infectious Diseases Laboratory, Royal Children's Hospital, Brisbane, Queensland 4029, Australia Queensland Children's Medical Research Institute, Royal Children's Hospital, The University of Queensland, Brisbane, Queensland 4029, Australia.
Public Health Microbiology, Communicable Disease, Queensland Health Forensic and Scientific Services, Archerfield, Queensland, Australia.
出版信息
J Antimicrob Chemother. 2014 Jun;69(6):1526-32. doi: 10.1093/jac/dkt544. Epub 2014 Jan 26.
OBJECTIVES
Neisseria gonorrhoeae antimicrobial resistance (AMR) is a global problem heightened by emerging resistance to ceftriaxone. Appropriate molecular typing methods are important for understanding the emergence and spread of N. gonorrhoeae AMR. We report on the development, validation and testing of a Sequenom MassARRAY iPLEX method for multilocus sequence typing (MLST)-style genotyping of N. gonorrhoeae isolates.
METHODS
An iPLEX MassARRAY method (iPLEX14SNP) was developed targeting 14 informative gonococcal single nucleotide polymorphisms (SNPs) previously shown to predict MLST types. The method was initially validated using 24 N. gonorrhoeae control isolates and was then applied to 397 test isolates collected throughout Queensland, Australia in the first half of 2012.
RESULTS
The iPLEX14SNP method provided 100% accuracy for the control isolates, correctly identifying all 14 SNPs for all 24 isolates (336/336). For the 397 test isolates, the iPLEX14SNP assigned results for 5461 of the possible 5558 SNPs (SNP call rate 98.25%), with complete 14 SNP profiles obtained for 364 isolates. Based on the complete SNP profile data, there were 49 different sequence types identified in Queensland, with 11 of the 49 SNP profiles accounting for the majority (n = 280; 77%) of isolates. AMR was dominated by several geographically clustered sequence types. Using the iPLEX14SNP method, up to 384 isolates could be tested within 1 working day for less than Aus$10 per isolate.
CONCLUSIONS
The iPLEX14SNP offers an accurate and high-throughput method for the MLST-style genotyping of N. gonorrhoeae and may prove particularly useful for large-scale studies investigating the emergence and spread of gonococcal AMR.
目的
淋病奈瑟菌对抗微生物药物的耐药性（AMR）是一个全球性问题，而头孢曲松耐药性的出现则使这一问题更加严重。适当的分子分型方法对于了解淋病奈瑟菌 AMR 的出现和传播非常重要。我们报告了一种新的Sequenom MassARRAY iPLEX 方法的开发、验证和测试，该方法用于淋病奈瑟菌分离株的多位点序列分型（MLST）式基因分型。
方法
我们开发了一种 iPLEX MassARRAY 方法（iPLEX14SNP），该方法针对先前显示可预测 MLST 型的 14 个淋病奈瑟菌信息性单核苷酸多态性（SNP）进行靶向检测。该方法最初使用 24 个淋病奈瑟菌对照分离株进行验证，然后应用于 2012 年上半年在澳大利亚昆士兰州收集的 397 个测试分离株。
结果
iPLEX14SNP 方法对对照分离株的准确率为 100%，正确识别了所有 24 个分离株的 14 个 SNP（336/336）。对于 397 个测试分离株，iPLEX14SNP 对 5558 个可能的 SNP 中的 5461 个进行了赋值（SNP 检出率为 98.25%），其中 364 个分离株获得了完整的 14 个 SNP 谱。基于完整的 SNP 谱数据，在昆士兰州发现了 49 种不同的序列类型，其中 11 种 SNP 谱占多数（n=280；77%）的分离株。AMR 主要由几个具有地理聚集性的序列类型主导。使用 iPLEX14SNP 方法，每天可对多达 384 个分离株进行检测，每个分离株的检测成本不到 10 澳元。
结论
iPLEX14SNP 提供了一种准确、高通量的淋病奈瑟菌 MLST 式基因分型方法，对于研究淋病奈瑟菌 AMR 的出现和传播的大规模研究可能特别有用。
Zotero 插件