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直接培养、增菌和聚合酶链反应检测肉鸡群屠宰时空肠弯曲菌和结肠弯曲菌的敏感性

Sensitivity of Direct Culture, Enrichment and PCR for Detection of Campylobacter jejuni and C. coli in Broiler Flocks at Slaughter.

作者信息

Rodgers J D, Simpkin E, Lee R, Clifton-Hadley F A, Vidal A B

机构信息

Department of Bacteriology, Animal and Plant Health Agency, Addlestone, Surrey, UK.

出版信息

Zoonoses Public Health. 2017 Jun;64(4):262-271. doi: 10.1111/zph.12306. Epub 2016 Oct 22.

Abstract

Broiler chicken flocks are a significant source of Campylobacter jejuni and Campylobacter coli that result in the major public health problem of campylobacteriosis. Accurate estimates of the prevalence of both C. coli and C. jejuni in flocks would enhance epidemiological understanding, risk assessment and control options. This study combined results from a panel of 10 detection tests (direct culture, enrichment and PCR) on caecal samples from flocks at slaughter. A parallel interpretation approach was used to determine the presence of Campylobacter spp. and for C. jejuni and C. coli individually. The sample was considered positive if at least one method detected the target and this interpretation was taken to represent a 'proxy gold standard' for detection in the absence of a gold standard reference test. The sensitivity of each individual method to detect Campylobacter spp., C. jejuni and C. coli was then estimated relative to the proxy gold standard. Enrichment in adapted Exeter broth (deficient in polymyxin B) with a resuscitation step was 100% sensitive, whilst direct culture on modified charcoal cefoperazone deoxycholate agar (mCCDA) was highly sensitive (97.9%). Enrichment methods using Preston broth and Bolton broth were significantly less sensitive. Enrichment in Exeter broth promoted the recovery of C. jejuni, whilst enrichment in Bolton broth favoured C. coli. A RT-PCR detection test could identify 80% of flocks that were co-colonised with both species. This study found that 76.3% (n = 127) of flocks were colonised with Campylobacter spp. The majority (95.9%) of Campylobacter-positive flocks were colonised with C. jejuni; however, approximately one-third of positive flocks were simultaneously colonised with both C. jejuni and C. coli. The findings highlight the impact of different detection methodologies on the accuracy of the estimated incidence of both C. jejuni and C. coli entering the abattoir within broiler flocks and the associated public health risks.

摘要

肉鸡群是空肠弯曲菌和结肠弯曲菌的重要来源,这两种细菌会导致弯曲菌病这一重大公共卫生问题。准确估计鸡群中结肠弯曲菌和空肠弯曲菌的流行率,将有助于增强对流行病学的理解、风险评估和控制措施。本研究综合了对屠宰时鸡群盲肠样本进行的10种检测试验(直接培养、增菌培养和PCR)的结果。采用平行解读方法来确定弯曲菌属的存在情况,以及分别确定空肠弯曲菌和结肠弯曲菌的存在情况。如果至少有一种方法检测到目标菌,则该样本被视为阳性,在没有金标准参考试验的情况下,这种解读被视为检测的“替代金标准”。然后相对于替代金标准,估计每种单独方法检测弯曲菌属、空肠弯曲菌和结肠弯曲菌的灵敏度。在含有复苏步骤的改良埃克塞特肉汤(不含多粘菌素B)中增菌培养的灵敏度为100%,而在改良炭头孢哌酮脱氧胆酸盐琼脂(mCCDA)上直接培养的灵敏度很高(97.9%)。使用普雷斯顿肉汤和博尔顿肉汤的增菌培养方法的灵敏度明显较低。在埃克塞特肉汤中增菌培养有利于空肠弯曲菌的回收,而在博尔顿肉汤中增菌培养则有利于结肠弯曲菌。一种逆转录PCR检测试验可以识别80%同时感染这两种菌的鸡群。本研究发现,76.3%(n = 127)的鸡群感染了弯曲菌属。大多数(95.9%)弯曲菌阳性鸡群感染的是空肠弯曲菌;然而,大约三分之一的阳性鸡群同时感染了空肠弯曲菌和结肠弯曲菌。这些发现突出了不同检测方法对估计进入屠宰场的肉鸡群中空肠弯曲菌和结肠弯曲菌发病率准确性的影响以及相关的公共卫生风险。

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