Identification of Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis by polymerase chain reaction
Keywords:
Haemophilus influenzae, Streptococcus pneumoniae, Neisseria meningitidis, polymerase chain reaction, bacterial, meningitis
Abstract
Introduction: Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis are the main human pathogens that cause meningitis.Objective: Primers omp2, lytA and crgA were evaluated with H. influenzae, S. pnumoniae and N. meningitidis DNA in a multiplex PCR, determining the sensitivity and the specificity of the technique.
Materials and methods: Primers for H. influenzae outer membrane protein (omp2, 1000 pb), S. pneumoniae autolysin (lytA, 395 pb) and N. meningitidis, contact regulated gene (crgA, 230 pb) were evaluated in a multiplex PCR, determining the sensitivity and the specificity of the technique.
Results: Reproducible results were obtained with 50 nM of each of the three primers and annealing temperature of 57oC in the multiplex PCR, obtaining a sensitivity of 12.5 fg for H. influenzae and S. pneumoniae and 3.12 fg for N. meningitidis. No cross reactions with other microorganisms agents of meningitis or related with the genera, appeared.
Conclusions: The results for sensitivity and specificity suggest that the evaluated primers can be used for the development of a PCR in a multiplex format to the identification of the three main
pathogens that cause meningitis.
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References
1. Zwijnenburg PJ, van der Poll T, Roord JJ, van Furth AM. Chemotactic factors in cerebrospinal fluid during bacterial meningitis. Infec Immun. 2006;74:1445-51.
2. Bartfield AA. Bacterial meningitis. Prim Care Update Ob Gyns. 2000;7:49-54.
3. Saez LX, McCracken GH. Bacterial meningitis in children. Lancet. 2003;361:2139-48.
4. Gutiérrez CE. infecciones por Haemophilus influenzae en la población pediátrica. Biomédica. 2001;21:369-88.
5. Dagan R, Englehad D, Piccar E, Engelhard D, Israeli Pediatric Bacteremia and Meningitis Group. Epidemiology of invasive childhood pneumococcal infections in Israel. JAMA. 1992;268:3328-32.
6. Weisfelt M, de Gans J, van der Poll T, ven de Beek D. Pneumococcal meningitis in adults: new approaches to management and prevention. Lancet Neurol. 2006;5:332-42.
7. Lee SO, Ryu SH, Park SJ, Ryu J, Woo JH, Kim YS. Meningococcal disease in the Republic of Korea army: incidence and serogroups determined by PCR. J Korean Med Sci. 2003;18:163-6.
8. Pathan N, Faust SN, Levin M. Pathophysiology of meningococcal meningitis and septicaemia. Arch Dis Child. 2003;88:601-7.
9. Gray L, Fedorko D. Laboratory diagnosis of bacterial meningitis. Clin Microbiol Rev. 1992;5:130-45.
10. Correa A, Echavarría E, Estrada S, Lopera T, Ortis LE, Uribe MV, et al. Taller sobre meningitis bacteriana aguda. Infectio. 2001;5:31-4.
11. Tunkel AR, Hartman BJ, Kaplan SL, Kaufman BA, Roos KL, Scheld WM, et al. Practice guidelines for the management of bacterial meningitis. Clin Infect Dis. 2004;39:1267-84.
12. Overturf GD. Defining bacterial meningitis and other infections of the central nervous system. Pediatr Crit Care Med. 2005;6(3 Suppl.):14-8.
13. Louie M, Louie L, Simor AE. The role of DNA amplification technology in the diagnosis of infectious diseases. CMAJ. 2000;163:301-9.
14. Pitcher D, Saunders N, Owen R. Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Letts Appl Microbiol. 1989;8:151-6.
15. Forbes K, Bruce K, Ball A, Penniington T. Variation in length and sequence of porin (omp2) alleles of noncapsulate Haemophilus influenzae. Mol Microbiol. 1992;6:2107-12.
16. 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. Diagn Microbiol Infect Dis. 2004;49:249-54.
17. Taha MK. Simultaneous approach for nonculture PCR based identification and serogroup prediction of N. meningitidis. J Clin Microbiol. 2000;38:855-7.
18. NCBI. BLAST. [Consultada: julio 13 de 2006]. Disponible en: http://www.ncbi.nlm.nih.gov/BLAST.
19. DNASTAR. Software for molecular biology [Consultada: julio 13 de 2006]. Disponible en: http://www.dnastar.com.
20. University of Helsinky. FastPCR © 1998-2007 (for Windows). [Consultada: julio 13 de 2006]. Disponible en: http://www.biocenter.helsinki.fi/bi/Programs/fastpcr.htm.
21. Falla TJ, Crook DW, Brophy LN, Maskell D, Kroll JS, Moxon ER. PCR for capsular typing of Haemophilus influenzae. J Clin Microbiol. 1994;32: 2382-6.
22. Hobson RP, Williams A, Rawal K, Pennington TH, Forbes KJ. Incidence and spread of Haemophilus influenzae on an Antarctic base determined using the polymerase chain reaction. Epidemiol Infect.
1994;114:93-103.
23. Uzuka R, Kawashima H, Hasegama D, Ioi H, Amaha M, Kashiwagi Y, et al. Rapid diagnosis of bacterial meningitis by using multiplex PCR and real time PCR. Pediatr Int. 2004;46:551-4.
24. 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. J Clin Microbiol. 2001;39:1553-8.
25. Chanteau S, Sidikou F, Djibo S, Moussa A, Mindadou, H, Boisier P. Scaling up of PCR-based surveillance of bacterial meningitis in the African meningitis belt: indisputable benefits of multiplex PCR assay in Niger. Trans R Soc Trop Med Hyg. 2006;100:677-80.
26. Schoske R, Vallone PM, Ruitberg CM, Butler JM. Multiplex PCR design strategy used for the simultaneous amplification of 10 Y chromosome short tandem repeat (STR) loci. Anal Bioanal Chem. 2003;375:333-43.
27. Henegariu O, Heerema NA, Dlouhy SR, Vance GM, Vogt PH. Multiplex PCR: critical parameters and stepby- step protocol. Biotechniques. 1997;23:504-11.
2. Bartfield AA. Bacterial meningitis. Prim Care Update Ob Gyns. 2000;7:49-54.
3. Saez LX, McCracken GH. Bacterial meningitis in children. Lancet. 2003;361:2139-48.
4. Gutiérrez CE. infecciones por Haemophilus influenzae en la población pediátrica. Biomédica. 2001;21:369-88.
5. Dagan R, Englehad D, Piccar E, Engelhard D, Israeli Pediatric Bacteremia and Meningitis Group. Epidemiology of invasive childhood pneumococcal infections in Israel. JAMA. 1992;268:3328-32.
6. Weisfelt M, de Gans J, van der Poll T, ven de Beek D. Pneumococcal meningitis in adults: new approaches to management and prevention. Lancet Neurol. 2006;5:332-42.
7. Lee SO, Ryu SH, Park SJ, Ryu J, Woo JH, Kim YS. Meningococcal disease in the Republic of Korea army: incidence and serogroups determined by PCR. J Korean Med Sci. 2003;18:163-6.
8. Pathan N, Faust SN, Levin M. Pathophysiology of meningococcal meningitis and septicaemia. Arch Dis Child. 2003;88:601-7.
9. Gray L, Fedorko D. Laboratory diagnosis of bacterial meningitis. Clin Microbiol Rev. 1992;5:130-45.
10. Correa A, Echavarría E, Estrada S, Lopera T, Ortis LE, Uribe MV, et al. Taller sobre meningitis bacteriana aguda. Infectio. 2001;5:31-4.
11. Tunkel AR, Hartman BJ, Kaplan SL, Kaufman BA, Roos KL, Scheld WM, et al. Practice guidelines for the management of bacterial meningitis. Clin Infect Dis. 2004;39:1267-84.
12. Overturf GD. Defining bacterial meningitis and other infections of the central nervous system. Pediatr Crit Care Med. 2005;6(3 Suppl.):14-8.
13. Louie M, Louie L, Simor AE. The role of DNA amplification technology in the diagnosis of infectious diseases. CMAJ. 2000;163:301-9.
14. Pitcher D, Saunders N, Owen R. Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Letts Appl Microbiol. 1989;8:151-6.
15. Forbes K, Bruce K, Ball A, Penniington T. Variation in length and sequence of porin (omp2) alleles of noncapsulate Haemophilus influenzae. Mol Microbiol. 1992;6:2107-12.
16. 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. Diagn Microbiol Infect Dis. 2004;49:249-54.
17. Taha MK. Simultaneous approach for nonculture PCR based identification and serogroup prediction of N. meningitidis. J Clin Microbiol. 2000;38:855-7.
18. NCBI. BLAST. [Consultada: julio 13 de 2006]. Disponible en: http://www.ncbi.nlm.nih.gov/BLAST.
19. DNASTAR. Software for molecular biology [Consultada: julio 13 de 2006]. Disponible en: http://www.dnastar.com.
20. University of Helsinky. FastPCR © 1998-2007 (for Windows). [Consultada: julio 13 de 2006]. Disponible en: http://www.biocenter.helsinki.fi/bi/Programs/fastpcr.htm.
21. Falla TJ, Crook DW, Brophy LN, Maskell D, Kroll JS, Moxon ER. PCR for capsular typing of Haemophilus influenzae. J Clin Microbiol. 1994;32: 2382-6.
22. Hobson RP, Williams A, Rawal K, Pennington TH, Forbes KJ. Incidence and spread of Haemophilus influenzae on an Antarctic base determined using the polymerase chain reaction. Epidemiol Infect.
1994;114:93-103.
23. Uzuka R, Kawashima H, Hasegama D, Ioi H, Amaha M, Kashiwagi Y, et al. Rapid diagnosis of bacterial meningitis by using multiplex PCR and real time PCR. Pediatr Int. 2004;46:551-4.
24. 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. J Clin Microbiol. 2001;39:1553-8.
25. Chanteau S, Sidikou F, Djibo S, Moussa A, Mindadou, H, Boisier P. Scaling up of PCR-based surveillance of bacterial meningitis in the African meningitis belt: indisputable benefits of multiplex PCR assay in Niger. Trans R Soc Trop Med Hyg. 2006;100:677-80.
26. Schoske R, Vallone PM, Ruitberg CM, Butler JM. Multiplex PCR design strategy used for the simultaneous amplification of 10 Y chromosome short tandem repeat (STR) loci. Anal Bioanal Chem. 2003;375:333-43.
27. Henegariu O, Heerema NA, Dlouhy SR, Vance GM, Vogt PH. Multiplex PCR: critical parameters and stepby- step protocol. Biotechniques. 1997;23:504-11.
How to Cite
1.
Parra E, Castañeda E, Moreno J. Identification of Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis by polymerase chain reaction. biomedica [Internet]. 2007 Sep. 1 [cited 2024 May 17];27(3):454-60. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/208
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