一种多重 PCR 检测方法，用于从坏死性筋膜炎患者的分离株中检测创伤弧菌、嗜水气单胞菌、耐甲氧西林金黄色葡萄球菌、化脓性链球菌和无乳链球菌。
A multiplex PCR assay for detection of Vibrio vulnificus, Aeromonas hydrophila, methicillin-resistant Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus agalactiae from the isolates of patients with necrotizing fasciitis.
机构信息
Department of Orthopedic Surgery, Chang Gung Memorial Hospital at Chia-Yi, Taiwan, Republic of China; College of Medicine, Chang Gung University at Taoyuan, Taiwan, Republic of China.
Department of Orthopedic Surgery, Chang Gung Memorial Hospital at Chia-Yi, Taiwan, Republic of China; College of Medicine, Chang Gung University at Taoyuan, Taiwan, Republic of China.
出版信息
Int J Infect Dis. 2019 Apr;81:73-80. doi: 10.1016/j.ijid.2019.01.037. Epub 2019 Jan 25.
BACKGROUND
Vibrio vulnificus, Aeromonas hydrophila, methicillin-resistant Staphylococcus aureus (MRSA), group A Streptococcus, and group B Streptococcus are commonly detected causative agents of necrotizing fasciitis (NF) in Chia-Yi Chang Gung Memorial Hospital. The aim of this study was to develop and evaluate a multiplex PCR method for the simultaneous detection of five of the most important human pathogens involved in NF by using a novel combination of species-specific genes.
METHODS
The samples used were collected from 99 patients with surgically confirmed NF of the extremities who were hospitalized consecutively between June 2015 and November 2017. Two sets of blood and tissue samples were collected from all patients; one set was sent to a microbiology laboratory for bacterial identification and the other set was sent to an immunohistochemistry laboratory for PCR amplification.
RESULTS
The multiplex PCR results for the blood samples showed negative findings. The multiplex PCR results for the tissue specimens showed 28 positive findings. Fourteen (87.5%) of the 16 V. vulnificus culture-positive tissue specimens, six (75%) of the eight A. hydrophila culture-positive tissue specimens, and four (100%) of the four MRSA culture-positive tissue specimens were positive by PCR. Similarly, two (100%) of the group A Streptococcus and two (100%) of the group B Streptococcus were PCR-positive.
CONCLUSIONS
The accuracy rate of the multiplex PCR presenting positive results in these culture-positive tissue samples was 87.5% (28/32). This suggests that multiplex PCR of tissue specimens may be a useful and rapid diagnostic tool for the detection of these lethal microorganisms in patients with NF.
背景
创伤弧菌、嗜水气单胞菌、耐甲氧西林金黄色葡萄球菌（MRSA）、A 组链球菌和 B 组链球菌是嘉义长庚纪念医院坏死性筋膜炎（NF）中常见的病原体。本研究旨在开发和评估一种多重 PCR 方法，使用一种新的种特异性基因组合，同时检测与 NF 相关的五种最重要的人类病原体。
方法
使用连续收集的 99 例四肢手术确诊的 NF 患者的样本。所有患者均采集两组血液和组织样本，一组送往微生物实验室进行细菌鉴定，另一组送往免疫组织化学实验室进行 PCR 扩增。
结果
血液样本的多重 PCR 结果为阴性。组织标本的多重 PCR 结果显示 28 个阳性发现。16 例创伤弧菌培养阳性的组织标本中，14 例（87.5%）通过 PCR 呈阳性；8 例嗜水气单胞菌培养阳性的组织标本中，6 例（75%）通过 PCR 呈阳性；4 例耐甲氧西林金黄色葡萄球菌培养阳性的组织标本中，4 例（100%）通过 PCR 呈阳性。同样，两组 A 组链球菌和两组 B 组链球菌均为 PCR 阳性。
结论
在这些培养阳性的组织样本中，多重 PCR 呈现阳性结果的准确率为 87.5%（28/32）。这表明，组织标本的多重 PCR 可能是一种有用且快速的诊断工具，可用于检测 NF 患者中的这些致死性微生物。
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