Multiplex polymerase chain reaction assay developed to diagnose adult bacterial meningitis in Taiwan.
Acute bacterial meningitis causes high morbidity and mortality; the associated clinical symptoms often are insensitive or non-specific; and the pathogenic bacteria are geographically diverse. Clinical diagnosis requires a rapid and accurate methodology. This study aimed to develop a new multiplex polymerase chain reaction (mPCR) assay to detect simultaneously six major bacteria that cause adult bacterial meningitis in Taiwan: Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus aureus, Escherichia coli, and Acinetobacter baumannii. Species-specific primers for the six bacteria were developed using reference strains. The specificities of the mPCRs for these bacteria were validated, and the sensitivities were evaluated via serial dilutions. The mPCR assay specifically detected all of the six pathogens, particularly with sensitivities of 12 colony forming units (CFU)/mL, 90 CFU/mL, and 390 CFU/mL for E. coli, S. pneumoniae, and K. pneumoniae, respectively. This mPCR assay is a rapid and specific tool to detect the six major bacterial pathogens that cause acute adult meningitis in Taiwan, particularly sensitive for detecting E. coli, S. pneumoniae, and K. pneumoniae. The assay may facilitate early diagnosis and guidance for antimicrobial therapy for adult patients with this deadly disease in Taiwan.
Department of Clinical Pathology, Buddhist Dalin Tzu Chi Hospital, Chiayi County, Taiwan.
Chi-Tsung
CT
Lee
Department of Life Science, National Chung Cheng University, Chiayi County, Taiwan.
Kuang-Ming
KM
Hsiao
Department of Neurosurgery, Buddhist Dalin Tzu Chi Hospital, Chiayi County, Taiwan.
Jin-Cherng
JC
Chen
Department of Clinical Pathology, Buddhist Dalin Tzu Chi Hospital, Chiayi County, Taiwan.|Anatomic Pathology, Buddhist Dalin Tzu Chi Hospital, Chiayi County, Taiwan.|Departments of Laboratory Medicine and Pathology, School of Medicine, Tzu Chi University, Hualien, Taiwan.
Cheng-Chuan
CC
Su
D040981:Acinetobacter baumannii / Q000235:genetics / Q000302:isolation & purification; D000328:Adult; D001431:Bacteriological Techniques / Q000379:methods / Q000706:statistics & numerical data; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D005798:Genes, Bacterial; D006801:Humans; D007711:Klebsiella pneumoniae / Q000235:genetics / Q000302:isolation & purification; D016920:Meningitis, Bacterial / Q000175:diagnosis* / Q000382:microbiology*; D020814:Meningitis, Escherichia coli / Q000175:diagnosis / Q000382:microbiology; D008586:Meningitis, Pneumococcal / Q000175:diagnosis / Q000382:microbiology; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000302:isolation & purification; D013624:Taiwan
D017931:DNA Primers; D004269:DNA, Bacterial
Adult; bacterial meningitis; cerebrospinal fluid; diagnosis; multiplex PCR