Sensitive and rapid detection of pathogenic bacteria from urine samples using multiplex recombinase polymerase amplification†,Lab on a Chip

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Sensitive and rapid detection of pathogenic bacteria from urine samples using multiplex recombinase polymerase amplification†
Lab on a Chip ( IF 6.1 ) Pub Date : 2018-07-02 00:00:00 , DOI: 10.1039/c8lc00399h
Junge Chen _{1,

2,

3,

4,

5} , Youchun Xu _{1,

2,

3,

4,

5} , He Yan _{1,

2,

3,

4,

5} , Yunzeng Zhu _{1,

2,

3,

4,

5} , Lei Wang _{5,

6,

7,

8,

9} , Yan Zhang _{5,

8,

9} , Ying Lu _{1,

2,

3,

4,

5} , Wanli Xing _{1,

2,

3,

4,

5}

Affiliation

Bacterial infections may cause severe diseases such as tuberculosis, sepsis, nephritis and cystitis. The rapid and sensitive detection of bacteria is a prerequisite for the treatment of these diseases. The current gold standard for bacterial identification is bacteriological culture. However, culture-based identification takes 3–7 days, which is time-consuming and laborious. In this study, bacteria in urine samples were enriched using a portable filter-based pipette. Then, a centrifugal chip was constructed to detect multiple pathogenic bacteria from urine samples by integrating the DNA extraction, multiplex recombinase polymerase amplification (RPA) and fluorescent detection together. This eliminated the time-consuming cultivation step, and thus accelerated the diagnosis of the urinary tract infections (UTIs). The five major pathogenic bacteria in UTIs were detected in this study, which are Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus and Salmonella typhimurium. Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa and Staphylococcus aureus were successfully detected with limits of detection of 100 CFU mL⁻¹ from urine samples within 40 min. Salmonella typhimurium was successfully detected with a limit of detection of 1000 CFU mL⁻¹ from urine samples. The chip-based bacteria detection proposed in this study is a promising tool for sensitive, accurate, and multiplex identification of bacteria in clinical urine samples of UTIs and bacteriuria.

中文翻译：

使用多重重组酶聚合酶扩增灵敏，快速地检测尿液样本中的病原细菌†

细菌感染可能导致严重的疾病，例如肺结核，败血症，肾炎和膀胱炎。快速而灵敏地检测细菌是治疗这些疾病的先决条件。当前用于细菌鉴定的金标准是细菌培养。但是，基于文化的识别需要3到7天，这既费时又费力。在这项研究中，使用便携式过滤器移液管富集了尿液样本中的细菌。然后，通过将DNA提取，多重重组酶聚合酶扩增（RPA）和荧光检测结合在一起，构建了一个离心芯片来检测尿液样本中的多种病原细菌。这消除了费时的培养步骤，从而加快了尿路感染（UTI）的诊断。大肠埃希菌，变形杆菌，铜绿假单胞菌，金黄色葡萄球菌和鼠伤寒沙门氏菌。在40分钟内从尿液样品中检测到100 CFU mL ^-1的检出限，成功检测出大肠杆菌，变形杆菌，铜绿假单胞菌和金黄色葡萄球菌。成功检测出鼠伤寒沙门氏菌，检出限为1000 CFU mL ^-1从尿液样本中提取。这项研究中提出的基于芯片的细菌检测是一种有希望的工具，可用于对尿路感染和细菌尿的临床尿液样本中的细菌进行灵敏，准确和多重鉴定。

更新日期：2018-07-02

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