Polymerase chain reaction (PCR)-based methods: Promising molecular tools in dentistry

Shahi, Shahriar; Vahed, Sepideh Zununi; Fathi, Nazanin; Sharifi, Simin

doi:10.1016/j.ijbiomac.2018.05.085

Cited by 32 publications

(18 citation statements)

References 148 publications

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“…There are 95 enzymes variants of this group that have been identified to date (Evans and Amyes, 2014). These enzymes are intrinsic and naturally exist in the chromosomal DNA of A. baumannii inherited into all strains (Shahi et al, 2018). This fact makes it a wonderful marker successfully applied for the detection of A. baumannii at the species level (Shahi et al, 2018;Ghaima et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

“…A comparison of isolates at the subspecies level is required by the application of molecular typing methods (Sadeghi et al, 2016). The use of molecular tools, especially the Polymerase Chain Reaction (PCR), has a great impact on simplifying and specificity for identification, characterization, and taxonomy of infectious agents (Shahi et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

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Molecular Study of Acinetobacter Baumannii Using 16s Rrna And Bla Oxa-51 Gene Isolated from Hospitals in Duhok-kurdistan Region, Iraq

Qader

2021

uod

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Background and objective: Acinetobacter baumannii is pathogenic and multiresistant bacteria that cause nosocomial infection. The aim of this study are diagnosis A. baumannii by using specific bla OXA-51 and 16SrRNA. Methods: out of 150 Acinetobacter species samples were collected from patients in Azadi teaching hospital, Duhok Emergency hospital; and burn Duhok cosmetic and burn hospital from May 2020 to August 2020. The samples were phenotypically diagnosed by bacteriological strategy. Out of 100 samples were selected and by molecular level confirmed as Acinetobacter spp. Results: isolates were revealed as small, pale, and late-lactose fermenter colonies on MacConkey agar appeared as a creamy, opaque, andnon-hemolytic colony on blood agar All suspected isolates, as A. baumannii, were growing at 44ºC. By using the genus-specific bla OXA-51 primer that produced 353 bp amplification band and 75 samples were diagnosed by PCR as A. baumannii by 16S rRNA as a specific primer and showed 150 bp amplicon. 10 samples out of the 75 resembling A. baumannii were identified as a multidrug resistant isolates by method of diffusion. Disc, A. baumannii isolates displayed a high resistance rate of 85% to azithromycin and 80% to imipenem. Moreover, amikacin, meropenem, and gentamycin, Trimethoprim-Sulfamethoxazole, and ciprofloxacin, norfloxacin showed a low level of efficacy against A. baumannii isolates with the resistant rate of 88%, 90%, 90%, and 93% respectively.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecular Study of Acinetobacter Baumannii Using 16s Rrna And Bla Oxa-51 Gene Isolated from Hospitals in Duhok-kurdistan Region, Iraq

Qader

2021

uod

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“…There are numerous genotypic and phenotypic methods currently in use for the detection of Tet(X)-producing E. coli and Acinetobacter spp. Genotypic detection using PCR allows high sensitivity and specificity, but high-throughput detection cannot be achieved due to the lack of universal primers for each gene subtype (Cavanaugh and Bathrick, 2018 ; Shahi et al, 2018 ). Multiplex real-time PCR can simultaneously detect multiple gene subtypes but is unable to identify unknown genes (Hawkins and Guest, 2017 ; Minkus et al, 2019 ).…”

Section: Discussionmentioning

confidence: 99%

Rapid Detection of High-Level Tigecycline Resistance in Tet(X)-Producing Escherichia coli and Acinetobacter spp. Based on MALDI-TOF MS

Cui

Zheng

Tang

et al. 2020

Front. Cell. Infect. Microbiol.

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The emergence and spread of the novel mobile Tet(X) tetracycline destructases confer high-level tigecycline and eravacycline resistance in Escherichia coli and Acinetobacter spp. and pose serious threats to human and animal health. Therefore, a rapid and robust Tet(X) detection assay was urgently needed to monitor the dissemination of tigecycline resistance. We developed a rapid and simple assay to detect Tet(X) producers in Gram-negative bacteria based on matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). This MALDI Tet(X) test was based on the inactivation of tigecycline by a Tet(X)-producing strain after a 3-h incubation of bacterial cultures with tigecycline. Culture supernatants were analyzed using MALDI-TOF MS to identify peaks corresponding to tigecycline (586 ± 0.2 m/z) and a tigecycline metabolite (602 ± 0.2 m/z). The results were calculated using the MS ratio [metabolite/(metabolite + tigecycline)]. The sensitivity of the MALDI Tet(X) test with all 216 test strains was 99.19%, and specificity was 100%. The test can be completed within 3 h. Overall, the MALDI Tet(X) test is an accurate, rapid, cost-effective method for the detection of Tet(X)-producing E. coli and Acinetobacter spp. by determining the unique peak of an oxygen-modified derivative of tigecycline.

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“…Reverse transcription PCR (RT-PCR) is a type of PCR methods that uses reverse transcriptase enzyme to convert RNA molecules to cDNA molecules. Then cDNA works as a template sequence for the PCR reaction [ 46 ]. Quantitative PCR determines a DNA molecule with the help of fluorescent dye or fluorophore-attached DNA probe such as TagMan [ 46 ].…”

Section: Current Detection Methods For Sars-cov-2mentioning

confidence: 99%

“…Then cDNA works as a template sequence for the PCR reaction [ 46 ]. Quantitative PCR determines a DNA molecule with the help of fluorescent dye or fluorophore-attached DNA probe such as TagMan [ 46 ]. A typical RT-PCR method includes four steps.…”

Section: Current Detection Methods For Sars-cov-2mentioning

confidence: 99%

Diagnostic techniques for COVID-19 and new developments

Sheikhzadeh

Eissa

Ismail

et al. 2020

Talanta

127

129

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COVID-19 pandemic is a serious global health issue today due to the rapid human to human transmission of SARS-CoV-2, a new type of coronavirus that causes fatal pneumonia. SARS -CoV-2 has a faster rate of transmission than other coronaviruses such as SARS and MERS and until now there are no approved specific drugs or vaccines for treatment. Thus, early diagnosis is crucial to prevent the extensive spread of the disease. The reverse transcription-polymerase chain reaction (RT-PCR) is the most routinely used method until now to detect SARS-CoV-2 infections. However, several other faster and accurate assays are being developed for the diagnosis of COVID-19 aiming to control the spread of infection through the identification of patients and immediate isolation. In this review, we will discuss the various detection methods of the SARS-CoV-2 virus including the recent developments in immunological assays, amplification techniques as well as biosensors.

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Polymerase chain reaction (PCR)-based methods: Promising molecular tools in dentistry

Cited by 32 publications

References 148 publications

Molecular Study of Acinetobacter Baumannii Using 16s Rrna And Bla Oxa-51 Gene Isolated from Hospitals in Duhok-kurdistan Region, Iraq

Molecular Study of Acinetobacter Baumannii Using 16s Rrna And Bla Oxa-51 Gene Isolated from Hospitals in Duhok-kurdistan Region, Iraq

Rapid Detection of High-Level Tigecycline Resistance in Tet(X)-Producing Escherichia coli and Acinetobacter spp. Based on MALDI-TOF MS

Diagnostic techniques for COVID-19 and new developments

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