|Author||Fukumoto H, Sato Y, Hasegawa H, Saeki H, Katano H.|
|Title||Development of a new real-time PCR system for simultaneous detection of bacteria and fungi in pathological samples.|
|Journal||Int J Clin Exp Pathol|
A novel system for simultaneous detection of pathogenic bacteria and fungi in pathological samples was developed using a real-time polymerase chain reaction (PCR) system. This system, designated the "multi-microbial real-time PCR", has the potential to simultaneously detect 68 bacterial and 9 fungal species in a 96-well plate format. All probe-primer sets were designed to produce amplicons smaller than 210 bp using formalin-fixed paraffin-embedded samples as input. The specificity and sensitivity of each probe-primer set were tested against DNA extracted from pure cultures of specific pathogens. The multi-microbial real-time PCR system revealed profiles of microorganism infection in lung samples collected at autopsy from 10 patients with acquired immunodeficiency syndrome. Staphylococcus aureus was the most common microbe detected (n=8), but with low copy numbers. High copy numbers of Pseudomonas aeruginosa were detected in the lung samples with abscess (n=6). Enterococcus faecium (n=6), Elizabethkingia meningoseptica (n=4), and Candida albicans (n=4) were also frequently detected. In addition, a latent infection of Mycobacterium tuberculosis was detected in one case of pneumonia. In conclusion, this multi-microbial real-time PCR system can be useful for detecting bacteria and fungi in pathological specimens from patients with uncertain diagnoses.
|MeSH||Adult Autopsy Bacteriological Techniques* DNA, Bacterial / classification DNA, Bacterial / genetics* DNA, Bacterial / isolation & purification DNA, Fungal / classification DNA, Fungal / genetics* DNA, Fungal / isolation & purification Female Humans Lung / microbiology* Male Middle Aged Multiplex Polymerase Chain Reaction* Predictive Value of Tests Real-Time Polymerase Chain Reaction* Reproducibility of Results Respiratory Tract Infections / diagnosis* Respiratory Tract Infections / microbiology|
|WOS Category||PATHOLOGY ONCOLOGY|
|DNA material||Genomic DNA of Staphylococcus aureus subsp. aureus JCM 2151 (JGD07701) Genomic DNA of Streptococcus pyogenes JCM 5674T (JGD12769) Genomic DNA of Streptococcus agalactiae JCM 5671T (JGD12768) Genomic DNA of Streptococcus sanguinis JCM 5708T (JGD07891) Genomic DNA of Clostridium difficile JCM 1296T (JGD12760) Genomic DNA of Peptostreptococcus anaerobius JCM 6477 (JGD12770) Genomic DNA of Actinomyces israelii JCM 12964T (JGD12771) Genomic DNA of Propionibacterium acnes JCM 6425T (JGD07462) Genomic DNA of Corynebacterium diphtheriae JCM 1310 (JGD07892) Genomic DNA of Nocardia asteroides JCM 3384T (JGD07465) Genomic DNA of Bacteroides fragilis JCM 11019T (JGD06283) Genomic DNA of Elizabethkingia meningoseptica JCM 21065T (JGD07973) Genomic DNA of Campylobacter jejuni subsp. jejuni JCM 2013 (JGD08023) Genomic DNA of Helicobacter pylori JCM 12095 (JGD07471) Genomic DNA of Stenotrophomonas maltophilia JCM 1975T (JGD12766) Genomic DNA of Citrobacter freundii JCM 1657T (JGD12761) Genomic DNA of Proteus mirabilis JCM 1669T (JGD12762) Genomic DNA of Morganella morganii subsp. morganii JCM 1672T (JGD12763) Genomic DNA of Providencia rettgeri JCM 1675T (JGD12764) Genomic DNA of Aspergillus niger JCM 22282 (JGD12767) Genomic DNA of Trichosporon asahii JCM 1809 (JGD12765).|