Simultaneous single-tube PCR assay for the detection of Neisseria meningitidis, Haemophilus influenzae type b and Streptococcus pneumoniae.
Rapid, accurate and inexpensive diagnosis of bacterial meningitis is critical for patient management. This study describes the development and evaluation of a multiplex PCR assay for the detection of Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae type b, which globally account for 90% of cases of bacterial meningitis. The single-tube assay, based on the ctrA, ply and bex targets, respectively, enabled detection of 5-10 pg DNA. When the assay was tested with clinical samples (n = 425), its sensitivity for the three targets was 93.9%, 92.3% and 88%, respectively, while the overall specificity and positive predictive value of the assay was 100%. The negative predictive value was 99.1-99.5%. The methodology permits rapid and accurate detection of the three main pathogens that cause bacterial meningitis.
Tzanakaki
G
G
National Meningococcal Reference Laboratory, National School of Public Health, Athens, Greece. gtzanakako@nsph.gr
Tsopanomichalou
M
M
Kesanopoulos
K
K
Matzourani
R
R
Sioumala
M
M
Tabaki
A
A
Kremastinou
J
J
D001426:Bacterial Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D004268:DNA-Binding Proteins / Q000235:genetics; D003937:Diagnosis, Differential; D019741:Haemophilus influenzae type b / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics; D016920:Meningitis, Bacterial / Q000175:diagnosis*; D008583:Meningitis, Haemophilus / Q000175:diagnosis; D008585:Meningitis, Meningococcal / Q000175:diagnosis; D008586:Meningitis, Pneumococcal / Q000175:diagnosis; D009345:Neisseria meningitidis / Q000235:genetics / Q000302:isolation & purification*; D011008:Pneumococcal Infections / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000302:isolation & purification*; D013301:Streptolysins / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics
D001426:Bacterial Proteins; C097706:CtrA protein, Caulobacter; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D026901:Membrane Transport Proteins; D013301:Streptolysins; D014157:Transcription Factors; C032290:plY protein, Streptococcus pneumoniae; C068102:bexB protein, Haemophilus influenzae