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通过分子生物学方法(赛沛GeneXpert IL、GeneOhm BD、罗氏LightCycler、Hyplex Evigene I2A)检测耐甲氧西林金黄色葡萄球菌(MRSA)与培养筛查的比较:实验室的经济实用策略

[Detection of Staphylococcus aureus resistant to methicillin (MRSA) by molecular biology (Cepheid GeneXpert IL, GeneOhm BD, Roche LightCycler, Hyplex Evigene I2A) versus screening by culture: Economic and practical strategy for the laboratory].

作者信息

Laudat P, Demondion E, Jouannet C, Charron J, Chillou C, Salaun V, Mankikian B

机构信息

Laboratoire Arnaud, 40, rue Jules-Simon, 37000 Tours, France.

出版信息

Pathol Biol (Paris). 2012 Jun;60(3):208-13. doi: 10.1016/j.patbio.2011.05.003. Epub 2011 Jul 5.

Abstract

OBJECTIVES

Patients admitted in cardiac surgery and cardiac ICU at the Clinic Saint-Gatien (Tours) are screened for MRSA at the entrance by nasal swab and culture on blood agar and selective chromogenic medium made by addition of cefoxitin: BBL CHROMagar MRSA-II BD (result obtained at Day +1). We wanted to assess the molecular biology techniques available to obtain a result at day 0 for the majority of patients and to define an economic and practical strategy for the laboratory.

TECHNIQUES

We studied four molecular biology techniques: Cepheid GeneXpert (Cepheid) GeneOhm (BD), LightCycler (Roche) and Hyplex (I2A). Upon reception, nasal swabs were treated by culture, considered as reference, and one of the techniques of molecular biology, according to the manufacturer's notice. We conducted four studies between April 2008 and February 2009 to obtain a significant sample for each of them.

METHODS

By screening we mean a method that allows us to exclude MRSA carriage for patients waiting for surgery, and not to change patient management: for example, lack of isolation measures specific to entrance, no modification of antibiotic prophylaxis during surgery and no isolation measures in the immediate postoperative period.

RESULTS

The criteria we considered for this evaluation were: (1) technician time: time to perform one or a series of sample(s) n=10 or more (about 2h for all techniques except GeneXpert 75min), level of skilled competences (no specific training for GeneXpert); (2) results: turnaround time (all molecular biology techniques), ease of reading and results interpretations (no specialized training required for GeneXpert), failure or not (12% of failure of internal controls for GeneOhm); (3) economic: cost for one or a series of sample(s) (n=10 or more), if we considered X as the reference culture cost (10 X Hyplex and LightCycler, 20 X and 40 X for GeneXpert GeneOhm); (4) NPV: 100% for GeneXpert and LightCycler.

CONCLUSION

At same sensitivity, no technique, including culture, can solve alone our problem, which is: (1) get results at day 0 for batch of samples (n<10): all molecular biology techniques; (2) beyond 10 samples: LightCycler (Roche) automated or Hyplex (I2A) manual; (3) when the result at day 1 is sufficient, the use of chromogenic agar with a reading of less than 18h as BBL CHROMagar MRSA II (BD) remains the most economical; (4) to be sure that a patient admitted at Day 0, even at night's emergency, is not carrier of MRSA: only Cepheid GeneXpert technology (IL). Furthermore, Cepheid GeneXpert (IL) allows performing several tests in parallel. The rapidity of this system can help control the transmission and make better use of antibiotics.

摘要

目的

在圣加蒂安诊所(图尔)心脏外科和心脏重症监护病房住院的患者,入院时通过鼻拭子在血琼脂和添加头孢西丁制成的选择性显色培养基(BBL CHROMagar MRSA-II BD)上进行培养来筛查耐甲氧西林金黄色葡萄球菌(MRSA)(第1天获得结果)。我们希望评估可用于在第0天为大多数患者获得结果的分子生物学技术,并为实验室确定一种经济实用的策略。

技术

我们研究了四种分子生物学技术:赛沛GeneXpert(赛沛)、GeneOhm(BD)、LightCycler(罗氏)和Hyplex(I2A)。收到鼻拭子后,按照制造商说明,通过培养(视为参考)和其中一种分子生物学技术进行处理。我们在2008年4月至2009年2月期间进行了四项研究,以便每项研究都能获得大量样本。

方法

我们所说的筛查是指一种方法,该方法能让我们排除等待手术患者的MRSA携带情况,且不改变患者管理方式:例如,入院时不采取特定隔离措施,手术期间不改变抗生素预防方案,术后即刻不采取隔离措施。

结果

我们用于此次评估的标准包括:(1)技术人员时间:处理一个或一系列样本(n = 10或更多)所需时间(除GeneXpert为75分钟外,所有技术约需2小时),技术能力水平(GeneXpert无需特殊培训);(2)结果:周转时间(所有分子生物学技术)、读数和结果解读的难易程度(GeneXpert无需专门培训)、是否失败(GeneOhm内部对照失败率为12%);(3)经济性:一个或一系列样本(n = 10或更多)的成本,如果我们将X视为参考培养成本(Hyplex和LightCycler为10X,GeneXpert和GeneOhm分别为20X和40X);(4)阴性预测值:GeneXpert和LightCycler均为100%。

结论

在相同敏感性下,没有任何一种技术(包括培养)能单独解决我们的问题,即:(1)对于一批样本(n < 10)在第0天获得结果:所有分子生物学技术;(2)超过10个样本:LightCycler(罗氏)自动化技术或Hyplex(I2A)手动技术;(3)当第1天的结果足够时,使用读数时间少于18小时的显色琼脂(如BBL CHROMagar MRSA II(BD))仍然是最经济的;(4)要确保第0天入院的患者(即使是夜间急诊)不是MRSA携带者:只有赛沛GeneXpert技术(IL)可行。此外,赛沛GeneXpert(IL)允许并行进行多项检测。该系统的快速性有助于控制传播并更好地使用抗生素。

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