Real-time PCR detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae DNA in clinical specimens


Authors: Z. Vacková 1,2;  D. Lžičařová 1,3;  N. K. Stock 1,4,5;  J. Kozáková 1
Authors‘ workplace: Centrum epidemiologie a mikrobiologie, Státní zdravotní ústav, Praha 1;  3. lékařská fakulta Univerzity Karlovy v Praze 2;  Ústav lékařské mikrobiologie 2. LF UK a FN Motol 3;  European Program for Public Health Microbiology (EUPHEM), ECDC, Stockholm, Sweden 4;  Institute of Hygiene and Microbiology, Julius-Maximilians-University, Würzburg, Germany 5
Published in: Epidemiol. Mikrobiol. Imunol. 64, 2015, č. 4, s. 222-230
Category: Original Papers

Overview

The study aim was to implement a molecular real-time polymerase chain reaction (PCR) assay recommended by the CDC (Centers for Disease Control and Prevention) for the detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical (culture negative) specimens from patients with suspected invasive bacterial disease. Clinical specimens are referred to the National Reference Laboratory (NRL) for Meningococcal Infections, Unit for Airborne Bacterial Infections, Centre for Epidemiology and Microbiology, National Institute of Public Health from various regions of the Czech Republic. Clinical specimens are, in particular, cerebrospinal fluid, anti-coagulated blood or serum and, exceptionally, post-mortem specimens. The NRL has implemented molecular diagnosis of these bacterial pathogens involved in meningitis and sepsis from clinical specimens since 1999. The first diagnostic method was semi-nested PCR followed by electrophoretic analysis. In 2014, a molecular qualitative real-time PCR assay was implemented.

Keywords:
rt-PCR detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae – rt-PCR ­detection of the agents of bacterial meningitis and sepsis


Sources

1. Kalmusová J, Pavlíková V, Křížová P, et al. První výsledky PCR diagnostiky invazivního meningokokového onemocnění. Klinická mikrobiologie a infekční lékařství, 2001;7(1):15–19. ISSN 1211-264X.

2. Kalmusová J, Bronská E, Křížová P. Diagnostika invazivního meningokokového, hemofilového a pneumokokového onemocnění PCR metodou. Klinická mikrobiologie a infekční lékařství, 2004;10:130–133. ISSN 1211-264X.

3. Carvalho MG, Tondella ML, McCaustland K, Weidlich L, McGee L, Mayer LW, Steigerwalt A, Whaley M, Facklam RR, Fields B, Carlone G, Ades EW, Dagan R, Sampson JS. Evaluation and improvement of real-time PCR assays targeting lytA, ply, and psaA genes for detection of pneumococcal DNA. Journal of Clinical Microbiology, 2007; 45(8):2460–2466. ISSN 0095-1137.

4. Messmer TO, Sampson JS, Stinson A, Wong B, Carlone GM, Facklam RR. Comparison of four polymerase chain reaction assays for specificity in the identification of Streptococcus pneumoniae. Diagnostic Microbiology and Infectious Disease, 2004;49:249–254. ISSN 0732-8893.

5. Mothershed EA, Sacchi CT, Whitney AM, Barnett GA, Ajello GW, Schmink S, Mayer LW, Phelan M, Taylor TH Jr, Bernhardt SA, Rosenstein NE, Popovic T. Use of real-time PCR to resolve slide agglutination discrepancies in serogroup identification of Neisseria meningitidis. Journal of Clinical Microbiology, 2004;42(1): 320–328. ISSN 0095-1137.

6. Taha MK, Alonso JM, Cafferkey M, Caugant DA, Clarke SC, Diggle MA, Fox A, Frosch M, Gray SJ, Guiver M, Heuberger S, Kalmusova J, Kesanopoulos K, Klem AM, Kriz P, Marsh J, Mölling P, Murphy K, Olcén P, Sanou O, Tzanakaki G, Vogel U. Interlaboratory comparison of PCR-based identification and genogrouping of Neisseria meningitidis. Journal of Clinical Microbiology, 2005;43(1):144–149. ISSN 0095-1137.

7. Wang X, Mair R, Hatcher C, Theodore MJ, Edmond K, Wu HM, Harcourt BH, Carvalho MG, Pimenta F, Nymadawa P, Altantsetseg D, Kirsch M, Satola SW, Cohn A, Messonnier NE, Mayer LW. Detection of bacterial pathogens in Mongolia meningitis surveillance with a new real-time PCR assay to detect Haemophilus influenzae. International Journal of Medical Microbiology, 2011;301:303–309. ISSN 1438-4221.

8. Corless CE, Guiver M, Borrow R, Edwards-Jones V, Fox AJ, Kaczmarski EB. Simultaneous Detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in Suspected Cases of Meningitis and Septicemia Using Real-Time PCR. Journal of Clinical Microbiology, 2001;39(4):1553–1558. ISSN 0095-1137.

9. WHO. PCR for Detection and Characterization of Bacterial Meningitis Pathogens: Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae, in Laboratory Methods for the Diagnosis of Meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae. WHO manual [online]. 2011, 2nd edition, Chapter 10. Dostupné na www: http://www.cdc.gov/meningitis/bacterial.html

10. CDC. Blood and Body Fluid DNA Extraction for Streptococci. CDC doporučení [online]. 2014, Dostupné na www: http://www.cdc.gov/streplab/downloads/pcr-body-fluid-DNA-extract-strep.pdf.

11. Emery SL, Erdman DD, Bowen MD, et al. Real-Time Reverse Transcriptiom-Polymerase Chain Reaction Assay for SARS-associated Coronavirus. Emerging Infectious Diseases, 2004;10(2):311–316. ISSN 1080-6040.

12. Piet JR, Huis in ’t Veld RAG, van Schaik BDC, et al. Genome Sequence of Neisseria meningitidis Serogroup B Strain H44/76. Journal of Bacteriology, 2011;193(9):2371–2372. ISSN 1098-5530.

13. Harrison A, Dyer DW, Gillaspy A, et al. Genomic Sequence of an Otitis Media Isolate of Nontypeable Haemophilus influenzae: Comparative Study with H. influenzae Serotype d, Strain KW20. Journal of Bacteriology, 2005;187(13): 4627–4636. ISSN 1098-5530.

14. Hiller NL, Janto B, Hogg JS, et al. Comparative Genomic Analyses of Seventeen Streptococcus pneumoniae Strains: Insights into the Pneumococcal Supragenome. Journal of Bacteriology, 2007;189(22):8186–8195. ISSN 1098-5530.

15. Hanage WP, Fraser C, Spratt BG. Fuzzy species among recombinogenic bacteria. BMC Biology, 2005;3:3–6. ISSN 1741-7007.

16. Dolan TJ, Hatcher CP, Satterfield DA, et al. sodC-Based Real-Time PCR for Detection of Neisseria meningitidis. PLoS ONE, 2011;6(5): e19361. ISSN 1932-6203.

17. Whatmore AW, Efstratiou A, Pickerill AP, et al. Genetic Relationships between Clinical Isolates of Streptococcus ­pneumoniae, Streptococcus oralis, and Streptococcus mitis: Characterization of “Atypical” Pneumococci and Organisms Allied to S. mitis Harboring S. pneumoniae Virulence Factor-Encoding Genes. Infection and Immunity, 2000;68(3):1374–1382. ISSN 1098-5522.

Labels
Hygiene and epidemiology Medical virology Clinical microbiology
Login
Forgotten password

Don‘t have an account?  Create new account

Forgotten password

Enter the email address that you registered with. We will send you instructions on how to set a new password.

Login

Don‘t have an account?  Create new account