Capsular type diversity of Mannheimia haemolytica determined by multiplex real-time PCR and indirect hemagglutination in clinical isolates from cattle, sheep, and goats in Spain

2021 ◽  
pp. 109121
Author(s):  
J.L. Arnal ◽  
A. Fernández ◽  
A.I. Vela ◽  
C. Sanz ◽  
J.F. Fernández-Garyzábal ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Mannheimia Haemolytica ◽  
Clinical Isolates ◽  
Capsular Type ◽  
Sheep And Goats ◽  
Indirect Hemagglutination

scholarly journals Real-Time PCR Assay for Detection of blaZ Genes in Staphylococcus aureus Clinical Isolates

2014 ◽  
Vol 52 (4) ◽  
pp. 1259-1261 ◽  
Author(s):  
L. A. Pereira ◽  
G. B. Harnett ◽  
M. M. Hodge ◽  
J. A. Cattell ◽  
D. J. Speers
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Clinical Isolates ◽  
Pcr Assay

scholarly journals Comparative evaluation of published cytomegalovirus primers for rapid real-time PCR: which are the most sensitive?

2009 ◽  
Vol 58 (7) ◽  
pp. 878-883 ◽  
Author(s):  
Wafa Habbal ◽  
Fawza Monem ◽  
Barbara C. Gärtner
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Analytical Sensitivity ◽  
Pcr Product ◽  
Highly Sensitive ◽  
Primer Sets ◽  
Analytical Sensitivities ◽  
Strain Ad169

Standardization of human cytomegalovirus (CMV) PCR is highly recommended. As primer design is essential for PCR sensitivity, this study evaluated all published CMV primer pairs to identify the most sensitive for single-round real-time PCR. PubMed (1993–2004) was searched for original papers aimed at CMV PCR. Fifty-seven papers were identified revealing 82 different primer pairs. Of these, 17 primer sets were selected for empirical study, as they were either used in real-time PCR or were evaluated comparatively by conventional PCR. After optimizing the PCR conditions, these primer sets were evaluated by real-time PCR using a SYBR Green format. Analytical sensitivities were assessed by testing the reference standard CMV strain AD169. A blast search was performed to identify mismatches with published sequences. Additionally, 60 clinical samples were tested with the three primer sets showing highest analytical sensitivity and the best match to all CMV strains. Three primer sets located in the glycoprotein B (UL55) gene region were found to be the most sensitive using strain AD169. However, two of these showed a considerable number of mismatches with clinical isolates in a blast search. Instead, two other pairs from the lower matrix phosphoprotein (UL83) gene and DNA polymerase (UL54) gene showed reasonable sensitivity and no mismatches with clinical isolates. These three pairs were further tested with clinical samples, which indicated that the two primer sets from UL55 and UL54 were the most sensitive. Interestingly, the analytical sensitivity of the PCR was inversely correlated with the size of the PCR product. In conclusion, these two primer pairs are recommended for a standardized, highly sensitive, real-time PCR.

Improved real-time PCR for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis clinical isolates

2003 ◽  
Vol 45 (3) ◽  
pp. 207-212 ◽  
Author(s):  
Maria J Torres ◽  
Antonio Criado ◽  
Maite Ruiz ◽  
Ana C Llanos ◽  
Jose C Palomares ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Clinical Isolates ◽  
Isoniazid Resistance

scholarly journals Use of Real-Time PCR and Fluorimetry for Rapid Detection of Rifampin and Isoniazid Resistance-Associated Mutations in Mycobacterium tuberculosis

2000 ◽  
Vol 38 (9) ◽  
pp. 3194-3199 ◽  
Author(s):  
Maria J. Torres ◽  
Antonio Criado ◽  
Jose C. Palomares ◽  
Javier Aznar
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Antimicrobial Agents ◽  
Dna Amplification ◽  
Single Strand Conformation Polymorphism ◽  
Clinical Isolates ◽  
Polymorphism Analysis ◽  
Antibiotic Resistant ◽  
Hybridization Probes

Very fast amplification of DNA in small volumes can be continuously monitored with a rapid cycler that incorporates fluorimetric detection. Primers were designed to amplify a 157-bp fragment of therpoB gene spanning codons 526 and 531 and a 209-bp fragment of the katG gene spanning codon 315 of Mycobacterium tuberculosis. Most mutations associated with resistance to rifampin (RMP) and isoniazid (INH) in clinical isolates occur in these codons. Two pairs of hybridization probes were synthesized; one in each pair was 3′ labeled with fluorescein and hybridized upstream of the codon with the mutation; the other two probes were 5′ labeled with LightCycler-Red 640. Each pair of probes recognized adjacent sequences in the amplicon. After DNA amplification was finished by using a LightCycler, the temperature at which the Red 640 probe melted from the product was determined in a 3-min melt program. Twenty M. tuberculosis clinical isolates susceptible to streptomycin, INH, RMP, and ethambutol and 36 antibiotic-resistant clinical M. tuberculosis isolates (16 resistant to RMP, 16 to INH, and 4 to both antimicrobial agents) were amplified, and the presence of mutations was determined using single-strand conformation polymorphism analysis, the LiQor automated sequencer, and the LightCycler system. Concordant results were obtained in all cases. Within 30 min, the LightCycler method correctly genotyped all the strains without the need of any post-PCR sample manipulation. Overall, this pilot study demonstrated that real-time PCR coupled to fluorescence detection is the fastest available method for the detection of RMP and INH resistance-associated mutations in M. tuberculosis clinical isolates.

Direct identification of Streptococcus agalactiae and capsular type by real-time PCR in vaginal swabs from pregnant women

2015 ◽  
Vol 21 (1) ◽  
pp. 34-38 ◽  
Author(s):  
Miyuki Morozumi ◽  
Naoko Chiba ◽  
Yuko Igarashi ◽  
Naoki Mitsuhashi ◽  
Takeaki Wajima ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Real Time ◽  
Streptococcus Agalactiae ◽  
Real Time Pcr ◽  
Capsular Type ◽  
Direct Identification ◽  
Vaginal Swabs

Detection of Malaysian methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates using simplex and duplex real-time PCR

2008 ◽  
Vol 25 (2) ◽  
pp. 253-258 ◽  
Author(s):  
Zarizal Suhaili ◽  
Saiful Azmi Johari ◽  
Mastura Mohtar ◽  
Ahmad Rushdi Tan Abdullah ◽  
Affandi Ahmad ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Clinical Isolates ◽  
Methicillin Resistant

scholarly journals Real-Time PCR and Statistical Analyses of acrAB and ramA Expression in Clinical Isolates of Klebsiella pneumoniae

2008 ◽  
Vol 52 (9) ◽  
pp. 3430-3432 ◽  
Author(s):  
Alexey Ruzin ◽  
Frederick W. Immermann ◽  
Patricia A. Bradford
Keyword(s):  
Klebsiella Pneumoniae ◽  
Real Time ◽  
Real Time Pcr ◽  
Statistical Analyses ◽  
Clinical Isolates ◽  
Susceptibility Breakpoint

ABSTRACT Clinical isolates of Klebsiella pneumoniae were tested for a correlation between tigecycline MIC and expression of ramA by using real-time PCR. At MICs of 4 and 8 μg/ml, the expression of ramA was statistically significantly different from MICs of 2 μg/ml or less, supporting the tigecycline susceptibility breakpoint of ≤2 μg/ml for K. pneumoniae.

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