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拭子类型和储存温度对 和 的分离和分子检测的影响。

Influences of swab types and storage temperatures on isolation and molecular detection of and .

机构信息

Institute of Infection and Global Health, University of Liverpool, Neston, UK.

Department of Veterinary Medical Sciences, University of Bologna, Ozzano dell'Emilia (BO), Italy.

出版信息

Avian Pathol. 2020 Feb;49(1):106-110. doi: 10.1080/03079457.2019.1675865. Epub 2019 Oct 22.

Abstract

Routine diagnosis of (MG) and (MS) is performed by collecting oropharyngeal swabs, followed by isolation and/or detection by molecular methods. The storage temperature, storage duration and the type of swab could be critical factors for successful isolation or molecular detection. The aim of this study was to compare the influence of different types of cotton-tipped swab stored at different temperatures, on the detection of MG and MS. To achieve this, combined use of traditional culture analysis (both agar and broth), with modern molecular detection methods was utilized. Performances of wooden and plastic shaft swabs, both without transport medium, were compared. Successful culture of was significantly more efficient from plastic swabs when compared to wooden, whereas no difference was seen for the re-isolation of . Storage at 4°C compared to room temperature also increased the efficiency of culture detection for both Mycoplasma species. When stored at room temperature, PCR detection limits of both MG and MS were significantly lower for wooden compared to plastic swabs. The qPCR data showed similar detection limits for both swab types when stored at both temperatures. The results suggest that swabs with a plastic shaft are preferred for MG and MS detection by both culture and PCR. While a lower storage temperature (4°C) is optimal for culture recovery, it seems that both temperatures investigated here are adequate for molecular detection and it is the swab type which carries a greater influence.

摘要

(MG)和(MS)的常规诊断通过采集口咽拭子进行,然后通过分子方法进行分离和/或检测。储存温度、储存时间和拭子类型可能是成功分离或分子检测的关键因素。本研究的目的是比较不同类型的棉花拭子在不同温度下储存对 MG 和 MS 检测的影响。为此,采用了传统的培养分析(琼脂和肉汤)与现代分子检测方法相结合的方法。比较了无运输介质的木制和塑料轴拭子的性能。与木制拭子相比,塑料拭子对的成功培养明显更有效,而对的再分离则没有差异。与室温相比,4°C 储存也提高了两种支原体的培养检测效率。在室温下储存时,与塑料拭子相比,MG 和 MS 的 PCR 检测下限均显著降低。qPCR 数据显示,两种拭子类型在两种温度下储存时的检测下限相似。结果表明,塑料轴拭子更适合通过培养和 PCR 检测 MG 和 MS。虽然较低的储存温度(4°C)有利于培养物的恢复,但这里研究的两种温度似乎都足以进行分子检测,而且拭子类型的影响更大。

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