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流式鉴定牛奶样本中的革兰氏阳性和阴性菌

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发表于 2012-11-20 13:24:55 | 显示全部楼层 |阅读模式

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该文用流式鉴定牛奶样本中的革兰氏阳性和阴性菌,使用的试剂是生物素标记的小麦胚芽凝集素和吖啶橙。首先用4株革兰阴性菌和15株革兰阳性菌进行测试,利用ROC曲线分析建立其cut-off值,然后再对53例标本进行检测,取得了较好的敏感度和特异度。不知道在人类外周血标本中是否可以鉴定?

全文见二楼。

Gram-typing of mastitis bacteria in milk samples using flow cytometry.
J Dairy Sci. 2012 Nov 7. pii: S0022-0302(12)00822-3. doi: 10.3168/jds.2012-5813. [Epub ahead of print]
Langerhuus SN   , Ingvartsen KL   , Bennedsgaard TW   , Røntved CM   .
Department of Animal Science, Faculty of Science and Technology, Aarhus University, Blichers Allé 20, Tjele, Denmark. Electronic address: Sine.Langerhuus@agrsci.dk.

Abstract
Fast identification of pathogenic bacteria in milk samples from cows with clinical mastitis is central to proper treatment. In Denmark, time to bacterial diagnosis is typically 24 to 48 h when using traditional culturing methods. The PCR technique provides a faster and highly sensitive identification of bacterial pathogens, although shipment of samples to diagnostic laboratories delays treatment decisions. Due to the lack of fast on-site tests that can identify the causative pathogens, antibiotic treatments are often initiated before bacterial identification. The present study describes a flow cytometry-based method, which can detect and distinguish gram-negative and gram-positive bacteria in mastitis milk samples. The differentiation was based on bacterial fluorescence intensities upon labeling with biotin-conjugated wheat germ agglutinin and acridine orange. Initially 19 in-house bacterial cultures (4 gram-negative and 15 gram-positive strains) were analyzed, and biotin-conjugated wheat germ agglutinin and acridine orange florescence intensities were determined for gram-negative and gram-positive bacteria, respectively. Fluorescence cut-off values were established based on receiver operating characteristic curves for the 19 bacterial cultures. The method was then tested on 53 selected mastitis cases obtained from the department biobank (milk samples from 6 gram-negative and 47 gram-positive mastitis cases). Gram-negative bacteria in milk samples were detected with a sensitivity of 1 and a specificity of 0.74, when classification was based on the previously established cut-off values. However, when receiver operating characteristic curves were constructed for the 53 mastitis cases, results indicate that a sensitivity and specificity of 1 could be reached if cut-off values were reduced. This flow cytometry-based technique could potentially provide dairy farmers and attending veterinarians with on-site information on bacterial gram-type and prevent ineffective antimicrobial treatment in mastitis cases caused by gram-negative bacteria.
Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
PMID: 23141826 [PubMed - as supplied by publisher]

发表于 2014-5-17 06:50:40 | 显示全部楼层
上传一下原文,另外在网上也找到一篇同样是丹麦那边做的牛奶微生物流式细胞分析的文章,同样共享给大家学习一下
A Flow-Cytometric Gram-Staining Technique for Milk-Associated Bacteria
免费在线阅读:Click Here to Read Online
下载PDF文档: A Flow-Cytometric Gram-Staining Technique for Milk-Associated Bacteria.pdf (367.04 KB, 下载次数: 9)
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