scholarly journals Polymerase Chain Reaction (PCR) and How Temperature Modifies Nucleotide Pairings

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Alicia García Falgueras
Keyword(s):  
Polymerase Chain Reaction ◽  
Immune System ◽  
Medical Treatment ◽  
Molecular Technique ◽  
Clinical Approach ◽  
Chain Reaction ◽  
Heating And Cooling ◽  
Long Time ◽  
Specific Virus ◽  
Polymerase Chain

Polymerase Chain Reaction (PCR) is a molecular technique more suitable for accurate diagnoses because of its specificity in targeting and identifying specific pathogens. PCR technique is reliable to detect the genetic presence of the virus in the host since the very beginning of the infection, no matter how the immune system of the person might react. Moreover, the samples for the test contains the DNA information during quite a long time since the extraction. Programmable thermal operation cycler for controlling heating and cooling is the procedure to denaturation into two single strands, annealing and extension reactions of the DNA. The PCR is certainly having a clinical approach, because do not emphasized on how the immune system of the patient might reacts to the infection, only on whether the specific virus is present or not for the accurate diagnoses and subsequently proceed with an appropriated medical treatment to restore health.

PCR BY THERMAL CONVECTION

2004 ◽  
Vol 18 (16) ◽  
pp. 775-784 ◽  
Author(s):  
DIETER BRAUN
Keyword(s):  
Polymerase Chain Reaction ◽  
Thermal Convection ◽  
Dna Amplification ◽  
Reaction Vessel ◽  
Cold Region ◽  
Chain Reaction ◽  
Heating And Cooling ◽  
Highly Sensitive ◽  
Specific Amplification ◽  
Polymerase Chain

The Polymerase Chain Reaction (PCR) allows for highly sensitive and specific amplification of DNA. It is the backbone of many genetic experiments and tests. Recently, three labs independently uncovered a novel and simple way to perform a PCR reaction. Instead of repetitive heating and cooling, a temperature gradient across the reaction vessel drives thermal convection. By convection, the reaction liquid circulates between hot and cold regions of the chamber. The convection triggers DNA amplification as the DNA melts into two single strands in the hot region and replicates into twice the amount in the cold region. The amplification progresses exponentially as the convection moves on. We review the characteristics of the different approaches and show the benefits and prospects of the method.

scholarly journals Циклостойкие миниатюрные термоэлектрические модули

2019 ◽  
Vol 53 (5) ◽  
pp. 604
Author(s):  
М.П. Волков ◽  
И.А. Драбкин ◽  
Л.Б. Ершова ◽  
А.А. Назаренко
Keyword(s):  
Polymerase Chain Reaction ◽  
Test Data ◽  
Medical Equipment ◽  
Dna Analysis ◽  
Heat Fluxes ◽  
Chain Reaction ◽  
Periodic Heating ◽  
Thermoelectric Modules ◽  
Heating And Cooling ◽  
Polymerase Chain

AbstractIn the paper the test data on new cycle-resistant thermoelectric modules are presented and discussed. These modules can be applied in medical equipment for polymerase chain reaction (PCR) to carry out DNA analysis with the help of rapid periodic heating and cooling of biological probes. However, high density of heat fluxes and, as a result, significant mechanical stresses in miniature thermoelectric modules involve special requirements to their reliability. The company RMT Ltd. has developed a technology for the production of highly reliable miniature thermoelectric modules that allowed them to withstand more than 500 thousand heating-cooling cycles (from 20 to 100°C) with a rate of 20°C/s and more.

scholarly journals Molecular detection and occurrence of equine theileriosis in Arabian horses in Al-Najaf province/Iraq

2020 ◽  
Vol 57 (3) ◽  
pp. e166996
Author(s):  
Hayder Mohammad Al-Rammahi ◽  
Abdulameer Abed Hatem ◽  
Asaad Chasib Al-Atabi
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Molecular Detection ◽  
Molecular Technique ◽  
Chain Reaction ◽  
Blood Samples ◽  
Equine Piroplasmosis ◽  
Polymerase Chain

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the variations in the occurrences of equine piroplasmosis according to gender, age, and geographical areas.

Multiplex Nested Reverse Transcription-Polymerase Chain Reaction for Respiratory Viruses in Acute Otitis Media

2003 ◽  
Vol 112 (3) ◽  
pp. 252-257 ◽  
Author(s):  
Toshio Ishibashi ◽  
Hiroko Monobe ◽  
Masanobu Shinogami ◽  
Yuka Nomura ◽  
Jun Yano
Keyword(s):  
Polymerase Chain Reaction ◽  
Otitis Media ◽  
Reverse Transcription ◽  
Acute Otitis Media ◽  
Respiratory Viruses ◽  
Molecular Technique ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Polymerase Chain

Because respiratory viruses play an important role in the causation and pathogenesis of acute otitis media (AOM), determining which virus has infected a child is important with respect to vaccines and antiviral drugs. In some instances, this information might be used to prevent the occurrence of AOM. We used a rapid, economical, and sensitive diagnostic system involving a multiplex nested reverse transcription–polymerase chain reaction (RT-PCR) assay to detect various respiratory viruses in clinical specimens of middle ear fluid (MEF) from children with AOM in our hospital. Multiplex RT-PCR was completed on 40 MEF samples from 28 infants and children less than 6 years old with AOM. Viral RNA was detected in 17 MEF samples (43%). Respiratory syncytial virus type A was present in 12 samples, adenovirus in 3, rhinovirus in 2, and influenza A (H3N2) in 1. The multiplex RT-PCR assay is recommended to clinical laboratories that are considering adoption of a molecular technique for viral diagnosis.

scholarly journals Aerosol-generating procedures in thoracic surgery in the COVID-19 era in Malaysia

2020 ◽  
Vol 28 (8) ◽  
pp. 495-499
Author(s):  
Narasimman Sathiamurthy ◽  
Narendran Balasubbiah ◽  
Benedict Dharmaraj
Keyword(s):  
Polymerase Chain Reaction ◽  
Thoracic Surgery ◽  
Reverse Transcriptase ◽  
Personal Protective Equipment ◽  
Healthcare Workers ◽  
Polymerase Chain Reaction Method ◽  
Protective Equipment ◽  
Chain Reaction ◽  
Long Time ◽  
Polymerase Chain

Background The Covid-19 pandemic has caused changes in the surgical treatment of non-Covid patients, especially in thoracic surgery because most procedures are aerosol generating. Hospital Kuala Lumpur, where thoracic procedures are performed, was badly affected. We describe our experience in performing aerosol generating procedures safely in thoracic surgery during the Covid-19 era. Methods Medical records of patients who underwent thoracic surgery from March 18, 2020 to May 17, 2020 were reviewed retrospectively. All patients undergoing thoracic surgery were tested for Covid-19 using the reverse transcriptase polymerase chain reaction method. Patients with malignancy were observed for 10 to 14 days in the ward after testing negative. The healthcare workers donned personal protective equipment for all the cases, and the number of healthcare workers in the operating room was limited to the minimum required. Results A total of 44 procedures were performed in 26 thoracic surgeries. All of these procedures were classified as aerosol generating, and the mean duration of the surgery was 130 ± 43 minutes. None of the healthcare workers involved in the surgery were exposed or infected by Covid-19. Conclusion Covid-19 will be a threat for a long time and thoracic surgeons must continue to provide their services, despite having to deal with aerosol generating procedures, in the new normal. Covid-19 testing of all surgical candidates, using the reverse transcriptase polymerase chain reaction, donning full personal protective equipment for healthcare workers, and carefully planned procedures are among the measures suggested to prevent unnecessary Covid-19 exposure in thoracic surgery.

Polymerase chain reaction for the amplification of the 121-bp repetitive sequence ofSchistosoma mansoni: a highly sensitive potential diagnostic tool for areas of low endemicity

2014 ◽  
Vol 89 (6) ◽  
pp. 769-773 ◽  
Author(s):  
E. Ferrer ◽  
F. Pérez ◽  
I. Bello ◽  
A. Bolívar ◽  
M. Lares ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sensitivity And Specificity ◽  
Repetitive Sequence ◽  
Annealing Temperature ◽  
Immunological Methods ◽  
Molecular Technique ◽  
Analytical Sensitivity ◽  
Chain Reaction ◽  
Highly Sensitive ◽  
Polymerase Chain

AbstractSchistosomiasis is a disease caused by parasitic flatworms of the genusSchistosoma, whose diagnosis has limitations, such as the low sensitivity and specificity of parasitological and immunological methods, respectively. In the present study an alternative molecular technique requiring previous standardization was carried out using the polymerase chain reaction (PCR) for the amplification of a 121-bp highly repetitive sequence forSchistosoma mansoni.DNA was extracted from eggs ofS. mansoniby salting out. Different conditions were standardized for the PCR technique, including the concentration of reagents and the DNA template, annealing temperature and number of cycles, followed by the determination of the analytical sensitivity and specificity of the technique. Furthermore, the standardized PCR technique was employed in DNA extracted, using Chelex®100, from samples of sera of patients with an immunodiagnosis of schistosomiasis. The optimal conditions for the PCR were 2.5 mmMgCl2, 150 mmdeoxynucleoside triphosphates (dNTPs), 0.4 μmprimers, 0.75 U DNA polymerase, using 35 cycles and an annealing temperature of 63°C. The analytical sensitivity of the PCR was 10 attograms of DNA and the specificity was 100%. The DNA sequence was successfully detected in the sera of two patients, demonstrating schistosomiasis transmission, although low, in the community studied. The standardized PCR technique, using smaller amounts of reagents than in the original protocol, is highly sensitive and specific for the detection of DNA fromS. mansoniand could be an important tool for diagnosis in areas of low endemicity.

scholarly journals A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales (Delphinapterusleucas)

2017 ◽  
Author(s):  
Ming-An Tsai ◽  
I-Hua Chen ◽  
Jiann-Hsiung Wang ◽  
Shih-Jen Chou ◽  
Tsung-Hsien Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Immune System ◽  
Reverse Transcription ◽  
Reference Genes ◽  
Amplification Efficiency ◽  
Chain Reaction ◽  
Beluga Whales ◽  
Qrt Pcr ◽  
Polymerase Chain

Cytokines are fundamental for a functioning immune system, and thus, potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for detecting the expression of proteins against which monoclonal antibodies are not available. In this study, we developed a novel probe-based quantitative gene expression assay for immunological assessment of captive beluga whales ( Delphinapterusleucas ) that is one of the most common cetacean species on display in aquariums worldwide. Six immunologically relevant genes (IL-2Rα, -4, -10, -12, TNFα, and IFNγ) were selected for analysis, and two validated housekeeping genes (PGK1 and RPL4) with stable expression were used as reference genes. Sixteen blood samples were obtained from four animals with different health conditions and stored in RNAlater solution. These samples were used for RNA extraction followed by qRT-PCR analysis. Analysis of gene transcripts was performed by relative quantitation using the comparative Cq method with the integration of amplification efficiency and two reference genes. The expression levels of each gene in the samples from clinically healthy animals were normally distributed. Transcript outliers for IL-2Rα, IL-4, IL-12, TNFα, and IFNγ were noticed in four samples collected from two clinically unhealthy animals. This assay has the potential to identify immune system deviation from normal state, which is caused by health problems. Furthermore, knowing the immune status of captive cetaceans could help both trainers and veterinarians in implementing preventive approaches prior to disease onset.

scholarly journals A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales (Delphinapterusleucas)

2017 ◽  
Author(s):  
Ming-An Tsai ◽  
I-Hua Chen ◽  
Jiann-Hsiung Wang ◽  
Shih-Jen Chou ◽  
Tsung-Hsien Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Immune System ◽  
Reverse Transcription ◽  
Reference Genes ◽  
Amplification Efficiency ◽  
Chain Reaction ◽  
Beluga Whales ◽  
Qrt Pcr ◽  
Polymerase Chain

Cytokines are fundamental for a functioning immune system, and thus, potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for detecting the expression of proteins against which monoclonal antibodies are not available. In this study, we developed a novel probe-based quantitative gene expression assay for immunological assessment of captive beluga whales ( Delphinapterusleucas ) that is one of the most common cetacean species on display in aquariums worldwide. Six immunologically relevant genes (IL-2Rα, -4, -10, -12, TNFα, and IFNγ) were selected for analysis, and two validated housekeeping genes (PGK1 and RPL4) with stable expression were used as reference genes. Sixteen blood samples were obtained from four animals with different health conditions and stored in RNAlater solution. These samples were used for RNA extraction followed by qRT-PCR analysis. Analysis of gene transcripts was performed by relative quantitation using the comparative Cq method with the integration of amplification efficiency and two reference genes. The expression levels of each gene in the samples from clinically healthy animals were normally distributed. Transcript outliers for IL-2Rα, IL-4, IL-12, TNFα, and IFNγ were noticed in four samples collected from two clinically unhealthy animals. This assay has the potential to identify immune system deviation from normal state, which is caused by health problems. Furthermore, knowing the immune status of captive cetaceans could help both trainers and veterinarians in implementing preventive approaches prior to disease onset.

scholarly journals A View on Polymerase Chain Reaction as an Outstanding Molecular Diagnostic Technique in Periodontology

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Adileh Shirmohammadi ◽  
Amirreza Babaloo ◽  
Solmaz Maleki Dizaj ◽  
Farzaneh Lotfipour ◽  
Simin Sharifi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Periodontal Disease ◽  
English Language ◽  
Diagnostic Technique ◽  
Molecular Diagnostic ◽  
Molecular Technique ◽  
Chain Reaction ◽  
Effective Technique ◽  
Publication Time ◽  
Polymerase Chain

Objectives. This study presents a discussion on the fundamentals of polymerase chain reaction (PCR) and its use as a diagnostic tool in periodontology. Materials and Methods. A computer-aided as well as hand-made search in PubMed and Scopus indexed journals (relevant to the topic) was done by keywords of molecular technique in periodontology, PCR, applications of PCR, and PCR in periodontics. Only the papers in the English language and outlining PCR and its association with periodontology were collected and utilized to provide a succinct review. There was no limitation for publication time. Results. The results of our search showed that PCR has turned into a standard in diagnosis in the field of periodontology. A variety of researches has demonstrated that its sensitive, and specific characteristics make it a quick and effective technique of recognition, identification, and quantification of microorganisms. Identification of various immunoinflammatory markers at the mRNA expression level as well as ascertaining gene-related polymorphisms can also be performed. Conclusions. The mechanisms of periodontal disease can further become clarified using PCR. Clinical Relevance. PCR as a diagnostic method can play a main part in the validation of the clinical diagnosis of periodontal disease indicating the reason, pathogenesis, clinical steps, progress, and prognosis of the disease.

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