| 1. |
- Abdeldaim, Guma M. K., et al.
(författare)
-
Quantitative fucK gene polymerase chain reaction on sputum and nasopharyngeal secretions to detect Haemophilus influenzae pneumonia
- 2013
-
Ingår i: Diagnostic microbiology and infectious disease. - : Elsevier. - 0732-8893 .- 1879-0070. ; 76:2, s. 141-146
-
Tidskriftsartikel (refereegranskat)abstract
- A quantitative polymerase chain reaction (PCR) for the fucK gene was developed for specific detection of Haemophilus influenzae. The method was tested on sputum and nasopharyngeal aspirate (NPA) from 78 patients with community-acquired pneumonia (CAP). With a reference standard of sputum culture and/or serology against the patient's own nasopharyngeal isolate, H. influenzae etiology was detected in 20 patients. Compared with the reference standard, fucK PCR (using the detection limit 10(5) DNA copies/mL) on sputum and NPA showed a sensitivity of 95.0% (19/20) in both cases, and specificities of 87.9% (51/58) and 89.5% (52/58), respectively. In a receiver operating characteristic curve analysis, sputum fucK PCR was found to be significantly superior to sputum P6 PCR for detection of H. influenzae CAP. NPA fucK PCR was positive in 3 of 54 adult controls without respiratory symptoms. In conclusion, quantitative fucK real-time PCR provides a sensitive and specific identification of H. influenzae in respiratory secretions.
|
|
| 2. |
- Barisic, Ivan, et al.
(författare)
-
Multiplex detection of antibiotic resistance genes using padlock probes
- 2013
-
Ingår i: Diagnostic microbiology and infectious disease. - : Elsevier BV. - 0732-8893 .- 1879-0070. ; 77:2, s. 118-125
-
Tidskriftsartikel (refereegranskat)abstract
- The elucidation of resistance mechanisms is of central importance to providing and maintaining efficient medical treatment. However, molecular detection methods covering the complete set of resistance genes with a single test are still missing. Here, we present a novel 100-plex assay based on padlock probes in combination with a microarray that allows the simultaneous large-scale identification of highly diverse beta-lactamases. The specificity of the assay was performed using 70 clinical bacterial isolates, recovering 98% of the beta-lactamase nucleotide sequences present. Additionally, the sensitivity was evaluated with PCR products and genomic bacterial DNA, revealing a detection limit of 10(4) DNA copies per reaction when using PCR products as the template. Pre-amplification of genomic DNA in a 25-multiplex PCR further facilitated the detection of beta-lactamase genes in dilutions of 10(7) cells/mL. In summary, we present an efficient, highly specific, and highly sensitive multiplex detection method for any gene.
|
|
| 3. |
|
|
| 4. |
- Karah, Nabil, et al.
(författare)
-
Plasmid-mediated quinolone resistance determinants qnr and aac(6')-Ib-cr in Escherichia coli and Klebsiella spp. from Norway and Sweden
- 2010
-
Ingår i: Diagnostic microbiology and infectious disease. - : Elsevier BV. - 0732-8893 .- 1879-0070. ; 66:4, s. 425-431
-
Tidskriftsartikel (refereegranskat)abstract
- The prevalence of the plasmid-mediated quinolone resistance genes qnr and aac(6')-Ib-cr was investigated among clinical isolates of Escherichia coli and Klebsiella spp. selected from 2 collections of consecutive isolates collected in 2004 to 2005 in Norway (n = 2479) and Sweden (n = 2980) and 1 group of extended-spectrum beta-lactamase (ESBL)-producing isolates collected in 2003 in Norway (n = 71). A total of 414 isolates was selected for screening based on resistance to nalidixic acid and/or reduced susceptibility/resistance to ciprofloxacin. The prevalence of both qnr and aac(6')-Ib-cr was higher among the ESBL producers (9.1% and 52.3%, respectively) than in the consecutive isolates (1.1% and 3.2%, respectively). qnrS1 was detected in 6 isolates, whereas qnrB1 and qnrB7 were detected in 2 isolates. The genetic structure surrounding qnrS1 was similar to previously described structures. In 2 isolates, qnrS1 was located on conjugative IncN-type plasmids of approximately 140 kb.
|
|
| 5. |
|
|
| 6. |
- Nakonieczna, Joanna, et al.
(författare)
-
Detection of Helicobacter rodentium-like DNA in the liver tissue of patients with chronic liver diseases by polymerase chain reaction-denaturing gradient gel electrophoresis and DNA sequence analysis
- 2010
-
Ingår i: Diagnostic Microbiology and Infectious Disease. - : Elsevier BV. - 1879-0070 .- 0732-8893. ; 68:3, s. 201-207
-
Tidskriftsartikel (refereegranskat)abstract
- Many Helicobacter spp. were isolated from the stomach, intestinal tract, and liver of different animals and humans. The association between Helicobacter spp. and hepatobiliary diseases, including hepatocellular carcinoma, was thoroughly examined, indicating a potential role of the bacteria in the progression toward cancer. In our work, we screened 97 liver biopsies from patients with chronic liver diseases for the presence of Helicobacter spp. DNA. With the use of genus-specific polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) and subsequent sequencing, we found that the majority of Helicobacter spp. DNA detected was similar to Helicobacter rodentium DNA (71%). The DNA of other detected Helicobacter spp. was similar to Helicobacter pylori DNA. This is the first indication of H. rodentium-like DNA presence in human liver tissue. We also conclude that PCR DGGE is a useful screening method for assigning species designation and heterogeneity. (C) 2010 Elsevier Inc. All rights reserved.
|
|
| 7. |
|
|
| 8. |
|
|
| 9. |
|
|
| 10. |
|
|
