Establishment and Performance Evaluation of Multiplex PCR-Dipstick DNA Chromatography for Mycoplasma pneumoniae and Chlamydia pneumoniae Rapid Detection

• Published in: The Canadian journal of infectious diseases & medical microbiology 2023-09, Vol.2023, p.1-7
• Main Authors: Hu, Liuyang, Wang, Xiuri, Cao, Donglin, Cheng, Qiuchen, Li, Qiong
• Format: Article
• Language: English
• Subjects: Analysis; Antibodies; Assaying; Bacterial pneumonia; Biosensors; Causes of; Chlamydia; Chlamydia pneumoniae; Chromatography; Deoxyribonucleic acid; Diagnosis; DNA; DNA sequencing; Drug resistance; Genetic research; Health aspects; Immunoglobulin M; Infectious diseases; Medical research; Medicine, Experimental; Mixed infection; Multiplexing; Mycoplasma pneumoniae; Nucleotide sequencing; Pathogens; Performance evaluation; Pneumonia; Political aspects; Polymerase chain reaction; Respiratory tract; Respiratory tract infection; Respiratory tract infections; Sexually transmitted diseases; Statistics; STD; Streptococcus infections; Viral antibodies
• ISSN: 1712-9532; 1918-1493
• DOI: 10.1155/2023/6654504

Background and Objective. Mycoplasma pneumoniae (MP) and Chlamydia pneumoniae (CP) infections are becoming increasingly prominent in respiratory infections. This study established a rapid, simple, sensitive, accurate multiplex PCR-dipstick DNA chromatography assay for Mycoplasma pneumoniae and Chlamydia pneumoniae detection. Methods. Nasopharyngeal swab samples of 300 children with an acute respiratory tract infection were detected by a multiplex PCR-dipstick chromatography assay, and the results were compared with the DNA sequencing and serum IgM antibody assay. Results. A multiplex PCR-dipstick DNA assay can specifically detect Mycoplasma pneumoniae and Chlamydia pneumoniae and shows a good specificity, with a minimum detection limit of 10 CFU/mL, respectively. Using DNA sequencing results as the gold standard, the sensitivity, specificity, positive predictive value, and negative predictive value of the multiplex PCR-dipstick DNA chromatography assay for the diagnosis of Mycoplasma pneumoniae were 96.61%, 100%, 100%, and 99.18% respectively, and those of Chlamydia pneumoniae were 95.24%, 100%, 100%, and 99.64% respectively. There was no statistical significance MP and CP diagnosis by the multiplex PCR-dipstick DNA assay and DNA sequencing (MP: P=0.5; CP: P=1.0), and the two assays had very high statistical consistency (MP: kappa = 0.979; CP: kappa = 0.974). The positive rate of the multiplex PCR-dipstick chromatography assay was significantly higher than that of the serum IgM antibody assay, with MP (17.7% vs. 13.3%), CP (5.7% vs. 3.3%), and mixed infection of MP and CP (1.3% vs. 0.67%). Conclusions. A multiplex PCR-dipstick chromatography assay was successfully established for the joint detection of Mycoplasma pneumoniae and Chlamydia pneumoniae within 2 hours. It is simple, fast, sensitive, accurate, cost-effective with good diagnostic performance, which can be used for small laboratories and point-of-care diagnosis.