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采用融解温度图谱法对儿科临床标本中细菌病原体的检测的前瞻性研究。

Prospective Study of the Detection of Bacterial Pathogens in Pediatric Clinical Specimens Using the Melting Temperature Mapping Method.

机构信息

Division of Infectious Diseases and Immunology, Saitama Children's Medical Center, Saitama, Japan.

Department of Clinical Laboratory and Molecular Pathology, Faculty of Medicine, Academic Assembly, University of Toyama, Toyama, Japan.

出版信息

Microbiol Spectr. 2022 Jun 29;10(3):e0019822. doi: 10.1128/spectrum.00198-22. Epub 2022 Jun 8.

Abstract

The melting temperature () mapping method is a novel technique that uses seven primer sets without sequencing to detect dominant bacteria. This method can identify pathogenic bacteria in adults within 3 h of blood collection without using conventional culture methods. However, no studies have examined whether pathogenic bacteria can be detected in clinical specimens from pediatric patients with bacterial infections. Here, we designed a new primer set for commercial use, constructed a database with more bacterial species, and examined the agreement rate of bacterial species . Moreover, we investigated whether our system could detect pathogenic bacteria from pediatric patients using the mapping method and compared the detection rates of the mapping and culture methods. A total of 256 pediatric clinical specimens from 156 patients (94 males and 62 females; median age, 2 years [<18 years of age]) were used. The observed concordance rates between the mapping method and the culture method for both positive and negative samples were 76.4% (126/165) in blood samples and 79.1% (72/91) in other clinical specimens. The mapping detection rate was higher than that of culture using both blood and other clinical specimens. In addition, using the mapping method, we identified causative bacteria in pediatric clinical specimens quicker than when using blood cultures. Hence, the mapping method could be a useful adjunct for diagnosing bacterial infections in pediatric patients and may be valuable in antimicrobial stewardship for patients with bacterial infections, especially in culture-negative cases. This study provides novel insights regarding the use of the melting temperature () mapping method to identify the dominant bacteria in samples collected from pediatric patients. We designed a new set of primers for commercial use and developed a database of different bacteria that can be identified using these primers. We show that the mapping method could identify bacteria from blood samples and other clinical specimens. Moreover, we provide evidence that the mapping method has a higher detection rate than that of the culture-based methods and can achieve a relatively high agreement rate. We believe that our study makes a significant contribution to this field because rapid identification of the source of bacterial infections can drastically improve patient outcomes and impede the development of antibiotic-resistant bacteria.

摘要

熔解温度(Tm)图谱法是一种新颖的技术,它使用七组引物而无需测序来检测优势细菌。该方法可以在采血后 3 小时内识别成人的致病菌,而无需使用传统的培养方法。然而,尚无研究检测图谱法是否可用于检测患有细菌感染的儿科患者的临床标本中的致病菌。在这里,我们设计了一种新的商用引物组,构建了一个包含更多细菌物种的数据库,并检验了细菌种类的图谱法检测符合率。此外,我们还调查了我们的系统是否可以使用 Tm 图谱法从儿科患者的临床标本中检测到致病菌,并比较了 Tm 图谱法和培养法的检测率。共使用了 156 名患者(94 名男性,62 名女性;中位数年龄为 2 岁[<18 岁])的 256 份儿科临床标本。血液样本中 Tm 图谱法与培养法对阳性和阴性样本的观察一致性率分别为 76.4%(126/165),其他临床标本为 79.1%(72/91)。使用 Tm 图谱法检测的阳性率高于使用血液和其他临床标本培养法的阳性率。此外,使用 Tm 图谱法,我们可以比使用血液培养更快地鉴定儿科临床标本中的致病菌。因此,Tm 图谱法可能是诊断儿科患者细菌感染的有用辅助手段,对于细菌感染患者的抗菌药物管理可能具有重要价值,特别是在培养阴性的情况下。本研究提供了关于使用熔解温度(Tm)图谱法鉴定儿科患者样本中优势细菌的新见解。我们设计了一组新的商用引物,并开发了一个可以使用这些引物鉴定不同细菌的数据库。我们表明,Tm 图谱法可以鉴定血液样本和其他临床标本中的细菌。此外,我们提供了证据表明,Tm 图谱法的检测率高于培养法,且具有较高的一致性率。我们认为,我们的研究对该领域做出了重要贡献,因为快速鉴定细菌感染的来源可以极大地改善患者的预后,并阻止抗生素耐药细菌的产生。

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