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Journal of Clinical Microbiology
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Bacteriology

Loop-Mediated Isothermal Amplification Method Targeting the lytA Gene for Detection of Streptococcus pneumoniae

Mitsuko Seki, Yoshihisa Yamashita, Hirotaka Torigoe, Hiromasa Tsuda, Setsuko Sato, Masao Maeno
Mitsuko Seki
1Department of Oral Health Sciences
2Division of Functional Morphology, Dental Research Center, Nihon University School of Dentistry, Tokyo
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  • For correspondence: seki-m@dent.nihon-u.ac.jp
Yoshihisa Yamashita
3Department of Preventive Dentistry, Faculty of Dental Science, Kyushu University, Fukuoka, Japan
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Hirotaka Torigoe
1Department of Oral Health Sciences
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Hiromasa Tsuda
1Department of Oral Health Sciences
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Setsuko Sato
1Department of Oral Health Sciences
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Masao Maeno
1Department of Oral Health Sciences
2Division of Functional Morphology, Dental Research Center, Nihon University School of Dentistry, Tokyo
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DOI: 10.1128/JCM.43.4.1581-1586.2005
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ABSTRACT

It is difficult to separate Streptococcus pneumoniae from the genotypically similar species Streptococcus mitis and Streptococcus oralis, which are commensals of the human oral cavity. A novel nucleic acid amplification technique, loop-mediated isothermal amplification (LAMP), which amplifies DNA under isothermal conditions (63°C) with high specificity, efficiency, and rapidity, was examined regarding its applicability for detecting S. pneumoniae. An S. pneumoniae-specific LAMP primer targeting the lytA gene was designed. The primer specificity was validated using 10 Streptococcus and 7 non-Streptococcus species. Within 60 min, the assay could detect 10 or more copies of purified S. pneumoniae DNA with a sensitivity 1,000 times that of conventional PCR. Clinical isolates of 21 other strains (3 S. oralis, 17 S. mitis, and 1 Streptococcus species) that harbor virulence-factor-encoding genes (lytA or ply) were tried to differentiate S. pneumoniae. The detection of S. pneumoniae in clinical isolates was more selective using the LAMP method than using conventional PCR. Therefore, LAMP appears to be a sensitive and reliable means of diagnosing S. pneumoniae infection.

  • Copyright © 2005 American Society for Microbiology
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Loop-Mediated Isothermal Amplification Method Targeting the lytA Gene for Detection of Streptococcus pneumoniae
Mitsuko Seki, Yoshihisa Yamashita, Hirotaka Torigoe, Hiromasa Tsuda, Setsuko Sato, Masao Maeno
Journal of Clinical Microbiology Apr 2005, 43 (4) 1581-1586; DOI: 10.1128/JCM.43.4.1581-1586.2005

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Loop-Mediated Isothermal Amplification Method Targeting the lytA Gene for Detection of Streptococcus pneumoniae
Mitsuko Seki, Yoshihisa Yamashita, Hirotaka Torigoe, Hiromasa Tsuda, Setsuko Sato, Masao Maeno
Journal of Clinical Microbiology Apr 2005, 43 (4) 1581-1586; DOI: 10.1128/JCM.43.4.1581-1586.2005
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KEYWORDS

N-Acetylmuramoyl-L-alanine Amidase
Nucleic Acid Amplification Techniques
Pneumococcal Infections
Streptococcus pneumoniae

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