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通过额外的树脂处理提高角膜器官培养基微生物检测的灵敏度。

Increased sensitivity of microbiological testing of cornea organ culture medium by additional resin treatment.

作者信息

Skenderi Zemra, Giurgola Laura, Gatto Claudio, D'Amato Tóthová Jana, Pruß Axel, Schroeter Jan

机构信息

Institute for Transfusion Medicine, University Tissue Bank, Charité - Universitätsmedizin Berlin, Berlin, Germany.

Research and Development, Alchilife Srl, Ponte San Nicolo, Italy.

出版信息

BMJ Open Ophthalmol. 2018 Nov 10;3(1):e000173. doi: 10.1136/bmjophth-2018-000173. eCollection 2018.

Abstract

OBJECTIVE

This validation study investigates the treatment of cornea organ culture medium (Modified Eagle Medium, Biochrom GmbH, Berlin, Germany) with RESEP, a new medical device for antibiotics removal, before microbiological testing with BACTEC blood culture bottles.

METHODS AND ANALYSIS

10-100 colony forming units of and were inoculated in 9mL of cornea organ culture medium. In group A, the medium was withdrawn with RESEP and treated for 20 min at room temperature, and then inoculated in BACTEC Plus Aerobic/F/Anaerobic/F blood culture bottles. In group B, the medium, spiked by the inoculation of microorganism, was injected directly. For each strain, a growth control was performed, by direct inoculation of the microorganisms in BACTECvials (positive control). All samples were incubated in the automated BACTECblood culture system at 36°C ±1°C for maximum of 14 days or until a positive reading. The elimination of antibiotics from the medium by RESEP was determined by high-performance liqiud chromatography.

RESULTS

After 20 min of RESEP treatment, 100% (n=9) of streptomycin, 100% (n=9) of amphotericin B and 99.7% (n=9) of penicillin G were eliminated. In group A, all microorganisms were detected within 3 days of incubation with a sensitivity of 100% (n=99) and showed no significant delay compared with the positive controls. In group B, the overall sensitivity was 67.9% (n=96) with a significant delay until detection of microbial growth for all tested microorganisms except for .

CONLCLUSION

The use of RESEP to eliminate the antibiotics from cornea organ culture medium increases the sensitivity of the microbiological testing with BACTECPlus blood culture bottles significantly and fulfils the requirements of the European Pharmacopoeia method suitability test.

摘要

目的

本验证研究调查了在使用BACTEC血培养瓶进行微生物检测之前,使用新型抗生素去除医疗器械RESEP对角膜器官培养基(改良伊格尔培养基,德国柏林Biochrom GmbH公司)的处理效果。

方法与分析

将10 - 100个菌落形成单位的 和 接种于9mL角膜器官培养基中。A组中,用RESEP抽取培养基并在室温下处理20分钟,然后接种于BACTEC Plus需氧/F/厌氧/F血培养瓶中。B组中,直接注入接种了微生物的培养基。对于每种菌株,通过将微生物直接接种于BACTEC小瓶(阳性对照)进行生长对照。所有样品在自动BACTEC血培养系统中于36°C±1°C孵育最多14天或直至出现阳性读数。通过高效液相色谱法测定RESEP从培养基中去除抗生素的情况。

结果

RESEP处理20分钟后,链霉素的去除率为100%(n = 9),两性霉素B的去除率为100%(n = 9),青霉素G的去除率为99.7%(n = 9)。在A组中,所有微生物在孵育3天内均被检测到,灵敏度为100%(n = 99),与阳性对照相比无显著延迟。在B组中,总体灵敏度为67.9%(n = 96),除 外,所有测试微生物在检测到微生物生长之前均有显著延迟。

结论

使用RESEP从角膜器官培养基中去除抗生素可显著提高使用BACTEC Plus血培养瓶进行微生物检测的灵敏度,并符合欧洲药典方法适用性试验的要求。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2964/6243474/d87b46e5a289/bmjophth-2018-000173f01.jpg

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