Detection of Haemophilus influenzae by loop-mediated isothermal amplification (LAMP) of the outer membrane protein P6 gene.
It is difficult and time-consuming to distinguish Haemophilus influenzae from the genotypically similar Haemophilus parainfluenzae, which is a commensal of the human oral cavity. The novel nucleic acid amplification technique of loop-mediated isothermal amplification (LAMP), which amplifies DNA under isothermal conditions (63 degrees C) with high specificity, efficiency, and rapidity, was evaluated for H. influenzae detection. A H. influenzae-specific LAMP primer set was designed for the outer membrane protein P6 gene. Primer set specificity was validated using 4 Haemophilus spp. and 13 other species. Within 60 min, LAMP detected 100 or more copies of purified DNA with a sensitivity that was 10-fold higher than that of conventional PCR. This method can be used to differentiate H. influenzae from H. parainfluenzae strains. Thus, LAMP may represent a sensitive and reliable means of diagnosing H. influenzae infection.
Torigoe
Hirotaka
H
Department of Oral Health Sciences, Nihon University School of Dentistry, Tokyo 101-8310, Japan.
Seki
Mitsuko
M
Yamashita
Yoshihisa
Y
Sugaya
Atsuto
A
Maeno
Masao
M
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D006192:Haemophilus Infections / Q000382:microbiology*; D018073:Haemophilus Vaccines / Q000235:genetics*; D006193:Haemophilus influenzae / Q000235:genetics*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity
D001425:Bacterial Outer Membrane Proteins; D018073:Haemophilus Vaccines; C069990:P6 outer membrane protein, Haemophilus