Design of a multiplex PCR for Streptococcus pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae and Chlamydophila pneumoniae to be used on sputum samples
| Autor: | Kristoffer Strålin, Hans Holmberg, Hans Fredlund, Anders Bäckman, Per Olcén |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Adult
Microbiology (medical) Mycoplasma pneumoniae Adolescent Pilot Projects Mycoplasmataceae Biology medicine.disease_cause Polymerase Chain Reaction Sensitivity and Specificity Pathology and Forensic Medicine Microbiology Haemophilus influenzae Lower respiratory tract infection Streptococcus mitis Streptococcus pneumoniae medicine Animals Humans Immunology and Allergy False Positive Reactions Child False Negative Reactions Respiratory Tract Infections Aged Aged 80 and over Sputum General Medicine Chlamydophila pneumoniae Middle Aged medicine.disease biology.organism_classification Virology respiratory tract diseases medicine.symptom |
| Zdroj: | APMIS. 113:99-111 |
| ISSN: | 1600-0463 0903-4641 |
| DOI: | 10.1111/j.1600-0463.2005.apm1130203.x |
| Popis: | A multiplex PCR (mPCR) was developed for simultaneous detection of specific genes for Streptococcus pneumoniae (lytA), Mycoplasma pneumoniae (P1), Chlamydophila pneumoniae (ompA), and Haemophilus influenzae (16S rRNA, with verification PCR for P6). When the protocol was tested on 257 bacterial strains belonging to 37 different species, no false negatives and only one false positive were noted. One Streptococcus mitis out of thirty was positive for lytA. In a pilot application study of 81 sputum samples from different patients with suspected lower respiratory tract infection (LRTI), mPCR identified S. pneumoniae in 25 samples, H. influenzae in 29, M. pneumoniae in 3, and C. pneumoniae in 1. All samples culture positive for S. pneumoniae (n=15) and H. influenzae (n=15) were mPCR positive for the same bacteria. In a pilot control study with nasopharyngeal swabs and aspirates from 10 healthy adults, both culture and mPCR were negative. No PCR inhibition was found in any of the mPCR-negative sputum or nasopharyngeal samples. Whether all samples identified as positive by mPCR are truly positive in an aetiological perspective regarding LRTI remains to be evaluated in a well-defined patient material. In conclusion, the mPCR appears to be a promising tool in the aetiological diagnostics of LRTI. |
| Databáze: | OpenAIRE |
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