评估免疫磁珠分离检测法结合培养法以提高从环境样本中回收嗜肺军团菌1血清型的能力。

Evaluation of an immunomagnetic separation assay in combination with cultivation to improve Legionella pneumophila serogroup 1 recovery from environmental samples.

作者信息

Allegra S, Girardot F, Grattard F, Berthelot P, Helbig J H, Pozzetto B, Riffard S

机构信息

Université de Lyon; 42023 Université de Saint-Etienne; Groupe Immunité des Muqueuses et Agents Pathogènes, EA 3064, Faculté de Médecine,

出版信息

J Appl Microbiol. 2011 Apr;110(4):952-61. doi: 10.1111/j.1365-2672.2011.04955.x.

DOI:10.1111/j.1365-2672.2011.04955.x

PMID:21276145

Abstract

AIMS

Legionella isolation from environmental samples is often difficult because of the presence of heterotrophic-associated bacteria that frequently overgrow when using standard culture (ISO 11731, 1998; NF T90-431, 2003) methods. To improve Legionella pneumophila recovery from complex water samples (water from cooling towers, biofilms), we evaluated an immunomagnetic separation (IMS) assay using a monoclonal antibody raised against the lipopolysaccharide of Leg. pneumophila sg1 in combination with culture.

METHODS AND RESULTS

This study was conducted on 51 environmental specimens. The comparison between IMS-culture and standard culture (ISO 11731, 1998; NF T90-431, 2003) methods was made using ISO 17994, 2004 criteria for establishing equivalence between microbiological methods based on the upper and lower (XH and XL) values of the relative difference (95% confidence limit) and D as maximum acceptable deviation (value of the confidence limit).

CONCLUSIONS

We found that the average performance of IMS culture was higher than the reference method.

摘要

目的

由于存在异养相关细菌，使用标准培养方法（ISO 11731, 1998；NF T90 - 431, 2003）时这些细菌常常过度生长，所以从环境样本中分离嗜肺军团菌通常很困难。为了提高从复杂水样（冷却塔水、生物膜）中回收嗜肺军团菌的效率，我们评估了一种免疫磁珠分离（IMS）检测方法，该方法使用针对嗜肺军团菌血清1型脂多糖产生的单克隆抗体并结合培养法。

方法与结果

本研究对51份环境标本进行了检测。依据ISO 17994, 2004标准，基于相对差异（95%置信限）的上限和下限（XH和XL）以及作为最大可接受偏差的D（置信限值）来确定微生物学方法之间的等效性，从而对免疫磁珠分离 - 培养法和标准培养法（ISO 11731, 1998；NF T90 - 431, 2003）进行比较。