Gan Chi, Hu Jinfeng, Cao Qing, Zhao Ruike, Li Yuchan, Wang Zhigang, Tao Yue, Mo Xi
The Laboratory of Pediatric Infectious Diseases, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China.
Department of Infectious Diseases, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China.
Ann Transl Med. 2020 Mar;8(5):203. doi: 10.21037/atm.2020.01.34.
Delays in the diagnosis of pediatric osteoarticular infections (OAIs) can cause associated acute complications or long-term morbidity. This study attempts to develop a multiplex PCR-based assay that can rapidly and accurately detect the main pathogens involved in pediatric OAIs, namely, methicillin-sensitive or methicillin-resistant and .
A set of four gene-specific primers suitable for use in a one-tube PCR assay was designed to detect four common pathogens involved in pediatric OAIs, namely, for methicillin-sensitive , and for methicillin-resistant for and for . The multiplex PCR was first evaluated with 39 isolated clinical strains and further with 41 specimens collected from patients suspected of having OAIs.
Specific primer pairs were successfully designed, and the targeted genes were simultaneously amplified. The product sizes in the assay for , , and were 233, 158, 336 and 109 bp, respectively. Evaluation of the multiplex PCR with 39 isolated clinical strains and 41 specimens revealed 100% sensitivity and 100% specificity. The limit of detection of the multiplex PCR assay was approximately 1×10 CFU at the bacterial cell level.
This newly developed multiplex PCR assay, without sequencing, enables a rapid and accurate diagnosis of the major bacterial species in children with OAIs and might serve as an additional diagnostic approach for urgent pathogen determination.
小儿骨关节炎感染（OAIs）诊断的延迟可导致相关急性并发症或长期发病。本研究试图开发一种基于多重PCR的检测方法，该方法能够快速准确地检测小儿OAIs中涉及的主要病原体，即甲氧西林敏感或耐甲氧西林的[病原体名称未完整给出]。
设计了一组适用于单管PCR检测的四种基因特异性引物，用于检测小儿OAIs中涉及的四种常见病原体，即用于甲氧西林敏感[病原体名称未完整给出]的[引物名称未完整给出]，用于耐甲氧西林[病原体名称未完整给出]的[引物名称未完整给出]，用于[病原体名称未完整给出]的[引物名称未完整给出]和用于[病原体名称未完整给出]的[引物名称未完整给出]。首先用39株分离的临床菌株对多重PCR进行评估，然后用从疑似患有OAIs的患者收集的41份标本进一步评估。
成功设计了特异性引物对，并同时扩增了靶基因。在检测[病原体名称未完整给出]、[病原体名称未完整给出]、[病原体名称未完整给出]和[病原体名称未完整给出]时，产物大小分别为233、158、336和109 bp。用39株分离的临床菌株和41份标本对多重PCR进行评估，结果显示灵敏度和特异性均为100%。多重PCR检测在细菌细胞水平的检测限约为1×10 CFU。
这种新开发的无需测序的多重PCR检测方法能够快速准确地诊断患有OAIs儿童的主要细菌种类，并且可能作为紧急病原体确定的一种额外诊断方法。
