Development and Evaluation of a Rapid Detection Technology of Streptococcus Pneumoniae Isolates from Specimen without Culture Based on LAMP and Bioinformatics

Abstract:

Streptococcus pneumoniae (S. pneumoniae) was one common pathogen that caused pneumonia, meningitis, otitis media, and sepsis. The traditional culture methods costed long time and omission often existed. A new method was developed based on Loop-mediated Isothermal Amplification (LAMP). A series of primers of autolysin (lytA) from S. pneumoniae were designed with specific primers soft, and set up LAMP system in detection of S. pneumoniae. Comparison of specificity and sensitivity results with culture and PCR method in 50 samples collected from patients with respiratory infections. The data demonstrated that the selected primers and reaction system in LAMP were respond to S. pneumoniae,not cross reacted to others bacteria. The detection of sensitivity to LAMP was 10² CFU/mL. All of clinical specimens in the test, 9 cases (18.0%) were confirmed to be positive as S. pneumoniae,whereas, only 4(8.0%) were diagnosed by culture method. The LAMP technology has a specific, simple and accurate characteristic in differentiation S. Pneumoniae from clinical specimen.

• Periodical: Applied Mechanics and Materials (Volume 618)
• Pages: 392-396
• DOI: https://doi.org/10.4028/www.scientific.net/AMM.618.392
• Online since: August 2014
• Authors: Yu Li, Xiao Mei Zhang, Jiao Jiao Liu, Wen Jing Yu, Ming Cheng Li*
• Keywords: Bioinformatics, Lamp, Streptococcus Pneumonia

References

[1] SeiM, YamashitaY, TorigoeH, et al. Loop-mediated isothemal amplification method targeting LytA gene for detection of Streptococcus pneumoniae. J Clin Microbiol, 2005, 43(4)： 1581–1586.

DOI: 10.1128/jcm.43.4.1581-1586.2005

[2] Notomit T, Okayama H, Masubuehi H, et al. Loop-mediated isothermal amplification of DNA. J Nucleic Acids Res, 2000, 28(12)：E63.

[3] Llull D, Lopez R, Garcia E. Characteristic signatures of the lytA gene provide a basis for rapid and reliable diagnosis of Streptococcus pneumoniae infections. J Clin Microbiol, 2006, 44(4)： 1250-1256.

DOI: 10.1128/jcm.44.4.1250-1256.2006

[4] Kim DW, Kilgore PE, Kim EJ, et al. Loop-mediated isothermal amplification assay to detect Haemophilus influenzae type b in cerebrospinal fluid. J Clin Microbiol, 20011, 49: 3621-3626.

DOI: 10.1128/jcm.00515-11

[5] Nandakumar R, Whiting J, Fouad AF. Identification of selected respiratory pathogens in endodontic infections. Oral Surg Oral Med, 2008, 106: 145-151.

DOI: 10.1016/j.tripleo.2008.01.036

[6] Jourdain S, Drèze PA, Vandeven J, et al. Smeesters PR. Sequential multiplex PCR assay for determining capsular serotypes of colonizing S. pneumoniae. BMC Infect Dis 2011; 11: 100.

DOI: 10.1186/1471-2334-11-100

[7] Pai R, Gertz RE, Beall B. Sequential multiplex PCR approach for determining capsular serotypes of Streptococcus pneumoniae isolates. J Clin Microbiol, 2006; 44: 124-31.

DOI: 10.1128/jcm.44.1.124-131.2006

[8] Notomi T H. Okayama H. Masubuchi T et al. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res, 2000, 28: e63.

DOI: 10.1093/nar/28.12.e63

[9] Farzanehkhah M, Hajihashemi B, Dolatyar A, et al. Real-time assay as tool for detection lytA gene in Streptococcus pneumoniae isolates. Cell J 2014; 16: 5.