Real-time PCR detection of Neisseria gonorrhoeae susceptibility to penicillin
Real-time PCR detection of Neisseria gonorrhoeae susceptibility to penicillin
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DOI：
10.1093/jac/dkw291
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发表时间：
2016-11-01
影响因子：
5.2
通讯作者：
Whiley, David M.
中科院分区：
文献类型：
--
作者：
Buckley, Cameron;Trembizki, Ella;Whiley, David M.
本研究的目的是建立淋球菌porB基因的实时荧光PCR检测方法（PorB-PCR），用于淋球菌对青霉素的敏感性预测。这补充了先前描述的用于检测青霉素酶产生N. PorB-PCR检测使用六种探针来表征PorB 1b类蛋白的位置101和102的各种氨基酸组合，包括WT G101/A102和突变体G101 K/A102 D、G101 K/A102 N和G101 K/A102 G序列，以及PorB 1a序列。这些序列预测青霉素敏感性的能力最初使用2307 N进行评估。表型敏感性数据可用的淋病分离株。然后将该测定应用于N.淋病阳性临床标本（n=70）。特异性通过检测肠道奈瑟菌菌株（n=75）和N.淋病阴性临床标本（n=171）。用PorB-PCR检测G101/A102和PorB 1a序列的淋球菌分离株共鉴定了78.4%（分别为61.2%和17.2%）的青霉素敏感分离株，特异性为97.4%和99.3%，阳性预测值为98.8%和98.9%，其中PPNG菌株被同时鉴定和排除。将PorB-PCR检测应用于N.淋病阳性临床标本。未观察到N.淋病阴性样本，并没有交叉反应，观察到与non-gonococcal species.When平行使用与先前描述的PPNG-PCR，PorB-PCR方法有可能促进使用青霉素淋病的个体化治疗。
The objective of this study was to develop a real-time PCR assay targeting the gonococcal porB gene (PorB-PCR) for predicting susceptibility of Neisseria gonorrhoeae to penicillin. This complements a previously described PCR assay for detecting penicillinase-producing N. gonorrhoeae (PPNG) developed by our laboratory (PPNG-PCR).The PorB-PCR assay was designed using six probes to characterize various combinations of amino acids at positions 101 and 102 of the PorB1b class protein, including the WT G101/A102 and mutant G101K/A102D, G101K/A102N and G101K/A102G sequences, as well as the PorB1a sequence. The ability of these sequences to predict penicillin susceptibility was initially assessed using 2307 N. gonorrhoeae isolates from throughout Australia for which phenotypic susceptibility data were available. The assay was then applied to N. gonorrhoeae-positive clinical specimens (n=70). Specificity was assessed by testing commensal Neisseria strains (n=75) and N. gonorrhoeae-negative clinical specimens (n=171).Testing of the 2307 N. gonorrhoeae isolates using PorB-PCR to detect G101/A102 and PorB1a sequences identified a total of 78.4% (61.2% and 17.2%, respectively) of penicillin-susceptible isolates with specificities of 97.4% and 99.3% and positive predictive values of 98.8% and 98.9%, where PPNG strains were simultaneously identified and excluded. Similar performance data were obtained when the PorB-PCR assay was applied to the N. gonorrhoeae-positive clinical specimens. No false-positive results were observed for the N. gonorrhoeae-negative samples and no cross-reactions were observed with the non-gonococcal species.When used in parallel with the previously described PPNG-PCR, the PorB-PCR approach has the potential to facilitate individualized treatment of gonorrhoea using penicillin.