Identification of the most common pathogenic bacteria in patients with suspected sepsis by multiplex PCR

Arabestani, Mohammad Reza; Fazzeli, Hossein; Esfahani, Bahram Nasr

doi:10.3855/jidc.3856

Cited by 22 publications

(12 citation statements)

References 32 publications

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“…The horizontal transmission of antimicrobial resistance genes via mobile elements, such as plasmids and transposons, plays an important role in the evolution and dissemination of multi-drug resistance [22,28]. …”

Section: Discussionmentioning

confidence: 99%

Detection of Integrons and Staphylococcal Cassette Chromosome mec Types in Clinical Methicillin-resistant Coagulase Negative Staphylococci Strains

Hajiahmadi

Ghale

Alikhani

et al. 2017

Osong Public Health Res Perspect

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ObjectivesIntegrons are thought to play an important role in the spread of antibiotic resistance. This study investigates class 1 and 2 integron-positive methicillin-resistant coagulase-negative staphylococci strains isolated in Iran and characterizes their patterns of antimicrobial resistance.MethodsHundred clinical isolates of coagulase-negative staphylococci were characterized for integron content and staphylococcal cassette chromosome mec (SCCmec) type.ResultsSixteen isolates carried class 1 (intI1) integrons and four isolates carried class 2 (intI2) integrons. One resistance gene array was identified among the class 1 integrons (aadA1 cassette). The distribution of SCCmec types in 50 methicillin-resistant coagulase-negative staphylococci strains showed that SCCmec types III and V dominated among the tested strains.ConclusionThis is the first report of methicillin-resistant coagulase-negative staphylococci strains that carry two mobile genetic elements, including class 1 and 2 integrons and SCCmec, in Iran.

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Section: Discussionmentioning

confidence: 99%

Detection of Integrons and Staphylococcal Cassette Chromosome mec Types in Clinical Methicillin-resistant Coagulase Negative Staphylococci Strains

Hajiahmadi

Ghale

Alikhani

et al. 2017

Osong Public Health Res Perspect

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“…A previously described set of primer pair was selected to be employed as the IC gene. The primer is specific to the chromosome X of Drosophila melanogaster (19). The resulting PCR product was cloned into a common TA vector.…”

Section: Preparation Of Internal Control (Ic)mentioning

confidence: 99%

In silico design and in vitro development of a highly accurate test to detect Brucella species

Safari¹,

Shahnazari²,

Mard-Soltani³

et al. 2019

Turk J Med Sci

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Background: Conventional methods of detecting Brucella spp. suffer from technical and biological complications. Besides, newly characterized species of the genus Brucella could be neglected by previously designed polymerase chain reaction (PCR) tests. Therefore, a more accurate PCR-based test seems to be imminently needed. Materials and methods:Blood samples were collected from 39 patients diagnosed with brucellosis and 25 healthy controls. Multiple sequence alignments (MSA) were performed on 500 Omp2-related protein and gene sequences. Thereafter, specific primers were designed and synthesized for the regions with highest conservancy. The collected samples were assessed by PCR test. To overcome the cross-reactivity issue, PCR thermal program was optimized regarding annealing time and temperature. Results:The MSA results indicated that the N terminus region of the Omp2 protein (DNA 5' end) is associated with highest conservancy. Primers with highest specificity were designed and synthesized. A two-step PCR reaction was successfully designed and optimized. The desirable bands were observed in clinical samples with high accuracy.Conclusion: It should be pointed out that using a precisely designed primer pair would bring about early infection detection, more success to detect all natural variants and higher cost-to-efficacy ratio in comparison to other detection methods.

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“…Amplification methods (e.g., polymerase chain reaction [PCR] have been used to amplify specific target regions in the microbial genome. For amplification based techniques it include broad range assays, Pathogen specific assays and multiplex PCR assays (Mancini et al,2010)..For broad range assays further Identification of microorganisms can be performed by PCR algorithms, taxon-specific oligonucleotide microarray, or sequencing amplification (Arabestani et al,2014).Broad-range PCR targets the 16S rRNA gene, a consensus gene that is present in all bacteria and consists of two regions -conserved and variable. The conserved regions are targeted by universal primers for detection of the presence of a microorganism; the variable regions are targeted by genus or species-specific primers (Woese,1987).…”

Section: Int J Adv Res 4(8) 2192-2202mentioning

confidence: 99%

Updating in Diagnosis of Blood Stream Infection in Immunocompromised Patients With Haematological Malignancies.

Shawky¹,

ElAttar²,

Ahmed³

2016

IJAR

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Introduction: Diagnosis of bloodstream infections using microbiological cultures suffers from low sensitivity and reporting delay. Advanced molecular techniques introduced in many laboratories provide rapid results and may show improvements in patient outcomes. This study aimed to evaluate the usefulness of a molecular technique, broad-range 16S rRNA PCR followed by Genusspecific multiplex PCR and Candida specific nested PCR for the diagnosis of bloodstream infections in immunocompromised patients with haematological malignancies, compared to automated blood culture .Methodology: Conventional broad-range PCR and candida specific nested PCR were performed , on EDTA blood and plasma samples collected from different patients with haematological malignancies complaining of febrile neutropenia; Positive cases by broad range PCR identified by genus specific multiplex PCR. results were compared with those of automated blood culture.Results: Though blood culture is regarded as the gold standard, Broad range PCR evaluation showed Sensitivity of 83.3% ,specificity of 86.8%, Positive predictive value of 66.7%, Negative predictive value of 94.3%and accuracy of 86.0% .Candida specific nested PCR evaluation showed Sensitivity of 100% ,specificity of 89.6%, Positive predictive value of 28.6% , Negative predictive value of 100%and accuracy of 90%. and the results of both techniques correlated well with those of blood culture. Conclusions: Molecular assays are promising method that can be used in the rapid diagnosis of bloodstream infections but they seem not to be sufficient to replace microbial cultures which must associate these techniques.

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Identification of the most common pathogenic bacteria in patients with suspected sepsis by multiplex PCR

Cited by 22 publications

References 32 publications

Detection of Integrons and Staphylococcal Cassette Chromosome mec Types in Clinical Methicillin-resistant Coagulase Negative Staphylococci Strains

Detection of Integrons and Staphylococcal Cassette Chromosome mec Types in Clinical Methicillin-resistant Coagulase Negative Staphylococci Strains

In silico design and in vitro development of a highly accurate test to detect Brucella species

Updating in Diagnosis of Blood Stream Infection in Immunocompromised Patients With Haematological Malignancies.

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