Polymer Chain Reaction Research Articles

Covalent Biosensing Polymer Chain Reaction Enabling Periphery Blood Testing to Predict Tumor Invasiveness with a Platelet Procancerous Protein.

Periphery blood testing is an attractive and relatively less invasive way of early cancer screening. In this work, based on the latest understanding of the pivotal role of platelets in promoting cancer invasion, a method for detecting a procancerous protein overexpressed both on platelets and in cancer cells is developed. As a kinase, the enzymatic activity, abundance, and self-phosphorylation of this protein are all important factors influencing its procancerous activity. To simultaneously determine these three important biochemical parameters, electrochemical control is called upon to connect or disconnect a polymer chain reaction (PCR) primer with a small-molecule synthetic probe, and with the target protein, in a target-specific manner. The resulting PCR signal amplification greatly improves the sensitivity of the design and also enables direct detection of the protein and its catalytic activity as well as its self-phosphorylation in clinical periphery blood samples from hepatocellular carcinoma (HCC) patients. This may point to future application of the proposed method in the early screening of HCC to assist its diagnosis and treatment.

Effects of biofertilizer on soil microbial diversity and antibiotic resistance genes

Spread of antibiotic resistance or the presence of antibiotic resistance genes (ARGs) in pathogens is a globally recognized threat to human health. Numerous studies have shown that application of organic fertilizers may increase the risk of ARGs, however, the risk of resistance genes associated with biofertilizers is largely unknown. To investigate whether biofertilizer application introduces ARGs to the soil, we used high-throughput quantitative polymerization chain reaction (HT-qPCR) to explore the effect of biofertilizer application over three years on soil ARGs in three orchards with different locations in China. Redundancy analysis showed specific and significant differences in the beta diversity of soil bacteria and fungi between treatments (fertilizer vs. no fertilizer). One-way ANOVA analysis revealed findings of the main driver of the significant difference in microbial community structure between fertilizer and control treatment was the change in soil properties following the application of biofertilizer. A total of 139 ARGs and 27 MGEs (mobile genetic elements), and 46 ARGs and 6 MGEs from 11 major taxa were detected in biofertilizer and soil samples, respectively. Only the samples from Guangxi had significant differences in the detected number of ARGs and MGEs between fertilization and control. Through structural equation modeling (SEM), we found that soil properties indirectly affected ARGs by shaping bacterial diversity, while bacterial abundance directly affected ARGs. Biofertilizer application did not significantly alter the relative abundance of ARGs in soil due to the complexity of the soil environment and competition between exogenous and native microorganisms. This study provided new insights into the spread of the antibiotic resistome of the soil through biofertilizer applications.

The Role of Genital Mycoplasmas in Preterm Labor.

Background:Growth of Mycoplasma in genital tract can cause problems such as infertility, pelvic inflammatory disease (PID), and preterm labor. This study was designed to evaluate the role of these bacteria in preterm labor among individuals in Gorgan city which is located in north of Iran.Methods:The study included 100 women with complaints of pain in preterm labor before 37 weeks of pregnancy (case group) and 100 women with term labor (control group) who were referred to Shahid Sayyad Shirazi Teaching Hospital in Gorgan city, north of Iran. Vaginal swabs, collected from all of these women, were evaluated for genital Mycoplasma sp. by molecular method using specific primers with polymerization chain reaction (PCR). The comparison of results was done by conducting X 2 and p<0.05 was considered significant.Results:Genital Mycoplasma was detected in 78 cases (39%) of 200 vaginal samples. Genital Mycoplasma colonization rates in the preterm and term samples were 60% and 18%, respectively, with relative risk of 2.05 (1.78–2.37) (p=0.001). The proportion of Ureaplasma parvum (44% and 15%), Ureaplasma urealyticum (11%, 3%), and Mycoplasma homins (5%, 0%) was significantly higher in women with preterm birth (PTB) than term labor. No cases of Mycoplasma genitalum were detected in this study.Conclusion:There is a significant relationship between presence of genital Mycoplasma in vaginal secretion and the risk of preterm labor.

Evaluation of patients with acute pancreatitis associated with SARS-CoV-2 (COVID-19); The importance of lipase/lymphocyte ratio in predicting mortality.

SARS-CoV-2 is the cause of a pandemic with high mortality. In the present study, the effects of the lipase/lymphocyte ratio on mortality were investigated in cases diagnosed with Covid-19 and acute pancreatitis. A total of 21 patients who were diagnosed with Covid-19 and acute pancreatitis, 34 patients who were not diagnosed with COVID-19 but diagnosed with acute pancreatitis, and 55 healthy control groups were divided into 3 groups and included in the study retrospectively. The patients who had positive RT-PCR (real‑time polymerized chain reaction) test results were included in the study. Complete blood count and biochemical values ​​of the patients were compared with those of the control group. When the data of the cases diagnosed with COVID-19 and acute pancreatitis were examined retrospectively, the amylase, lipase, lipase/lymphocyte ratio, and D-dimer levels were found to be significantly higher than in the control group (p < 0.01). In the ROC analysis, the amylase, lipase, and lipase/lymphocyte ratio had a high AUC (area under the curve) value (0.993 / 0.949 / 0.978, respectively). The lipase/lymphocyte ratio can be used in cases diagnosed with Covid-19 and acute pancreatitis to predict mortality (Tab. 3, Fig. 3, Ref. 23).

PZR with the help of markers in the cotton plant DNA

This article provides information on the mechanism of polymer chain reaction in plant DNK using markers. In addition, we provide detailed information on the sequence of PZR analysis using DNK samples and markers, and the examination of DNK samples by electrophoresis.

Amplified UCNPs-Mitoxantrone dihydrochloride fluorescence PCR sensor based on inner filter for ultrasensitive and rapid determination of Salmonella typhimurium

A fluorescence sensor composed of alendronic acid (ADA) @upconversion nanoparticles (UCNPs) and mitoxantrone dihydrochloride (MTX) combined with polymer chain reaction (PCR) was developed for rapid, specific and ultrasensitive determination of Salmonella typhimurium. MTX, which has a wide absorption band at 500–700 nm, can completely overlap the upconversion fluorescence (UCF) emission spectrum, leading to the reduction of UCF intensity. By intercalating MTX into DNA structure, the absorption intensity of MTX is obviously weak. Thus, the concentration of target can be quantified by response change of UCF intensity based on inner filter (IFE) between UCNPs and MTX coupled with PCR. The sensitivity of sensor for detection of Salmonella typhimurium is improved due to IFE and PCR amplification process, with a detection limit of 11 CFU/mL. In addition, the UCNPs-MTX PCR fluorescence sensor has been applied to sensing Salmonella typhimurium in chicken samples with satisfied results. The research work revealed a rapid and sensitive analytical method for determination of Salmonella typhimurium and provided an insight for the construction of PCR fluorescence sensor.

Ex vivo Antimicrobial Evaluation of Different Drugs Using RT-PCR Test

Introduction: Various medicaments are used in conjugation with mechanical cleaning and shaping to render root canals free of microbes. The different medicaments used tend to react among themselves sometimes resulting in deleterious products. For this reason use of alternative medicine, therapy is advocated.&#x0D; Aim: To evaluate the antimicrobial extent of different homeopathic drugs in comparison to the Ayurvedic and Allopathic medicines.&#x0D; Materials and Methods: A biofilm model of 8 mm dentine discs was prepared from single-rooted teeth, which were inoculated with E. faecalis for 21 days. 100 samples were divided into six groups Group I: Saline (negative Control), Group II: 2% Chlorhexidine Gluconate, Group III: Propolis, Group IV: RHUS Glabra, Group V: Zincum Oxydatum. The infected biofilm was then treated with the medicaments for seven days followed by testing to obtain threshold cycle (Ct) values using the Real-time polymerase chain reaction technique. DNA isolation was carried out and amplification was done using 16S rRNA gene sequence-based nested species-specific primers. DNA denaturation was done using DNA Thermocycler and a threshold cycle was obtained using Real-time Polymerize chain reaction technique. &#x0D; Results: The threshold cycle (Ct) values obtained were subjected to statistical analysis that revealed a significant difference between the groups and among all 2% Chlorhexidine Gluconate, being the highest (30.3460±0.02505) followed by Propolis (30.1365±0.02621) then RHUS Glabra (30.0865±0.02581) better than Zincum Oxydatum (29.8070±0.02319) and least being Saline (29.6380±0.02505).&#x0D; Conclusion: All the homeopathic medicines showed antimicrobial properties comparable to the allopathic and ayurvedic drugs. Among RHUS Glabra and Zincum Oxydatum, RHUS Glabra performed significantly better.

Significance of vascular endothelial growth factor (VEGFA)-1154 G/A gene polymorphism (rs1570360) in rheumatoid arthritis patients

Aim of the workTo evaluate the association of the vascular endothelial growth factor (VEGF) 1154G/A single nucleotide polymorphism (SNP) with the risk of RA and its possible relation to different disease parameters. Patients and methodsFifty RA patients and 50 matched healthy controls were enrolled in the study. Disease activity score (DAS28) was assessed. VEGF 1154G/A was measured byreal time polymerized chain reaction (RT-PCR) for patients and controls. ResultsThe patients mean age was 44.2 ± 12.1 years (25–64 years) and they were 41 females and 9 males with a disease duration of 5 ± 3.6 years and DAS28 of 5.88 ± 1.06. None of the patients were in remission or had low disease activity. The VEGF-1154 AA genotype and A allele were significantly higher in controls (54%, 56%) compared to the patients (10%, 23%)(p < 0.001). The GG, GA genotypes and G allele tended to be higher in patients (64%, 26% and 77%) compared to the control (42%, 4% and 44% respectively (p < 0.001). As regards anti-cyclic citrullinated peptide (anti-CCP) and rheumatoid factor (RF), AA genotype was significantly higher in seronegative (93.1% and 96.5%) patients compared to seropositive (6.9% and 3.5%; p = 0.002 and p = 0.005 respectively). There was no significant difference in the genotype according to gender (p = 0.45) or DAS28 (p = 0.78). No significant association was found between the SNP with the age (p = 0.48), disease duration (p = 0.58), ESR (p = 0.97) and CRP (p = 1). ConclusionsThe minor A allele of VEGFA1154G/A might act as a protective factor against RA and might also be related to a better disease prognosis.

New insights on severe clinical manifestations and deaths from visceral leishmaniasis in free-living crab-eating foxes (Cerdocyon thous) in Brazil

Visceral leishmaniasis (VL) is a zoonotic and severe neglected tropical disease, with worldwide distribution, that still cause many deaths among dogs and humans. Brazil is the country responsible for about 97% of the cases of leishmaniasis in the Americas and the disease is still considered a serious public health concern. Among wild mammalians, studies demonstrate the involvement of Cerdocyon thous in the biological cycle of Leishmania. Nevertheless, several authors consider the clinical manifestation of the disease to be rare or mild in free-living animals. Herein, we demonstrate the occurrence of severe clinical sings and deaths caused by VL in free-living crab-eating foxes. Three specimens of foxes collected from periurban areas were diagnosed with VL. The animals presented cutaneous, ophthalmological, gastrointestinal, locomotor and hematological alterations, and died after clinical progression. We identified the presence of anti-Leishmania antibodies by immunochromatographic test in all specimens. We also observed intra and extracellular amastigotes in skin cytology and lymph node aspirate. Furthermore, Leishmania infantum DNA was identified in all samples by the polymer chain reaction technique. Additionally, we performed blood count and stool parasitological tests and observed hematological disorders common to VL, such as anemia and lymphopenia. Taken together, our data demonstrate that VL can induce clinical complications and even cause death in C. thous and corroborate that this crab-eating fox is an adequate host for L. infantum.

Metformin treatment reverses high fat diet- induced non-alcoholic fatty liver diseases and dyslipidemia by stimulating multiple antioxidant and anti-inflammatory pathways

PurposeThis current study investigated the effect of metformin treatment on hepatic oxidative stress and inflammation associated with nonalcoholic fatty liver disease (NADLD) in high fat diet (HFD) fed rats. MethodWistar rats were fed with a HFD or laboratory chow diet for 8 weeks. Metformin was administered orally at a dose of 200 mg/kg. Body weight, food and water intake were recorded on daily basis. Oral glucose tolerance test (OGTT), biochemical analysis and histological examinations were conducted on plasma and tissue samples. Antioxidant and anti-inflammatory mRNA expression was analyzed using reverse transcription polymeric chain reaction (RT-PCR). ResultsMetformin treatment for 8 weeks prevented HFD-induced weight gain and decreased fat deposition in HFD fed rats. Biochemical analysis revealed that metformin treatment significantly attenuated nitro-oxidative stress markers malondialdehyde (MDA), advanced protein oxidation product (APOP), and excessive nitric oxide (NO) levels in the liver of HFD fed rats. Gene expression analysis demonestrated that metformin treatment was associated with an enhanced expression of antioxidant genes such as Nrf-2, HO-1, SOD and catalase in liver of HFD fed rats. Metformin treatment also found to modulate the expression of fat metabolizing and anti-inflammatory genes including PPAR--γ, C/EBP-α, SREBP1c, FAS, AMPK and GLUT-4. Consistent with the biochemical and gene expression data, the histopathological examination unveiled that metformin treatment attenuated inflammatory cells infiltration, steatosis, hepatocyte necrosis, collagen deposition, and fibrosis in the liver of HFD fed rats. ConclusionIn conclusion, this study suggests that metformin might be effective in the prevention and treatment of HFD-induced steatosis by reducing hepatic oxidative stress and inflammation in the liver.

Plasma circPTPN22 and its potential target microRNA-200a-3p as novel diagnostic biomarkers in systemic lupus erythematosus patients

BackgroundSystemic lupus erythematosus (SLE) is a chronic autoimmune disease with heterogeneous manifestations and intricate pathophysiology. Driven by its complicated diagnostic framework, an interest in innovative diagnostic approaches has been steadily growing. Circular RNAs (circRNAs) are novel class of noncoding RNAs involved in the development of various diseases including SLE. Aim of the workTo investigate circPTPN22, microRNA-200a-3p (miR-200a-3p) and interleukin-2 (IL-2) in SLE patients and study their diagnostic potential. Patients and methodsThe study included 45 patients and 45 control. The expression of circPTPN22 and miR-200a-3p were evaluated using quantitative reverse transcription polymerized chain reaction PCR. In addition, plasma IL-2 levels were determined by ELISA. The SLE disease activity index (SLEDAI) was assessed. ResultsThe patients were 42 females and 3 males, and their mean age was 33.6 ± 7.9 years and disease duration 11 ± 6.5 years. The expression of circPTPN22 and IL-2 were significantly lower in patients (0.33 ± 0.18 and 35.7 ± 22.02 pg/ml) compared to control (1 ± 0.1 and 147 ± 153.3 pg/ml respectively; p < 0.001). Meanwhile, the expression of miR-200a-3p was significantly higher in patients (4.59 ± 1.35 vs 1.11 ± 0.35; p < 0.001). A significant negative correlation was noticeable between circPTPN22 and miR-200a-3p expression. No correlation was found between IL-2 plasma levels and either circPTPN22 or miR-200a-3p and there was no correlation with clinical manifestations or SLEDAI. circPTPN22, miR-200a-3p and IL-2 could significantly differentiate patients and controls (p < 0.0001); with an area under the curve (AUC) of 0.99, 0.998 and 0.89 respectively. ConclusionsCircPTPN22 and miR-200a-3p are potential non-invasive diagnostic biomarkers for SLE, where circPTPN22 might function through sponging miR-200a-3p and interrupting its function.

Evaluating the Efficiency of Newly Formulated Pomade® and Ceftiofur Hydrochloride for Treating Foot Rot in Dairy Cattle

Background: Foot rot is an important contagious disease that causes economic loss in dairy cattle. Even though many antibiotic treatments have been tried on foot rot, very few information about new topical treatment method or product have been reported about the disease. The main objective of this paper is to evaluate an alternative new treatment for foot root disease in dairy cattle. Methods: Forty-one swap samples were collected from cattle’s feet which were further investigated under microscopic examination and polymerase chain reaction (PCR) for Dichelobacter nodosus and Fusobacterium necrophorum. Newly formulated pomade® and ceftiofur hydrochloride (Eficur®) applications were used in foot rot cases. Result: Polymerized chain reaction revealed D. nodosus in all the samples were as F. necrophorum was seen only in 22 (53.66%) samples. In this study, D. nodosus was considered as the primary agent involved in foot disease of cattle and F. necrophorum along with other bacterias were considered to be associated in the infection. The foot rot wounds formed in all cases (100%) in which D. nodosus, F. necrophorum and other bacteria were detected were healed along with tissue regeneration. As a result of treatment applications, a success rate of 93.33% was obtained in lameness resulting due to medium foot rot cases and 45.45% in severe acute lameness. The lameness recovery rate was found to be 80.48%.

Polymer Chain Reaction (PCR): Principle and Applications

The new, standard molecular biologic system for duplicating DNA enzymatically devoid of employing a living organism, like E. coli or yeast, represents polymerases chain reaction (PCR). This technology allows an exponential intensification of a minor quantity of DNA molecule several times. Analysis can be straightforward with more DNA available.&#x0D; A thermal heat cycler performs a polymerization chain reaction that involves repeated cycles of heating and cooling the reactant tubes at the desired temperature for each reaction step. A heated deck is positioned on the upper reaction tube to avoid evaporating the reaction mixture (normally volumes range from 15 to 100 l per tube), or an oil layer can be placed on a reaction mixture surface.&#x0D; The amplified DNA fragment is determined based on selecting primers in addition to the starting and end of the DNA fragment. The primers stand for short, artificial DNA stripes, no higher than fifty (typically 18-25bp) nucleotides have been based on a starting and ending of DNA fragment to be amplified. DNA-polymerase connects and starts a new DNA strand synthesis&#x0D; The PCR products can be visualized by dual foremost methods: (1) staining of the product of DNA amplified by a chemical dye like bromide ethidium, or (2) marking of fluorescent dyes (fluorophores) PCR primers or nucleotides before amplification of PCRs.&#x0D; PCR offers some benefits. First, it is a simple method of understanding and using and quick results. It has an extremely sensitive technology with the potential for sequencing, cloning, and analyzing millions or milliards of copies of a particular product.

(Invited) Ultrasensitive Detection of Nucleic Acid Molecules with Interferometric Plasmonic Microscopy

The detection of nucleic acid molecules plays important role in clinical diagnosis of diseases and fundamental biomedical research. The widely used polymerized chain reaction technique requires hours of amplification process, and is prone to aerosol pollution. Here, we presented the amplification-free digital sensing of nucleic acid molecules with the interferometric plasmonic microscopy (iPM). A sandwich assay was performed with the primary DNA probes coated on the plasmonic sensing surface and the secondary DNA probes tethered Au nanoparticles. Hybridization between DNA probes and single analyte molecules was monitored by the imaging of single Au nanoparticle with iPM system. By digitally counting the events of hybridization, the detection limit at the fM level could be achieved. Without amplification, this assay could be done within one hour. Furthermore, by analyzing the real-time binding behaviro during single hybridization event, non-specific bindings could be excluded from the specific ones, resulting in the improved specificity. This technique could be further improving by a careful design of the kinetic binding properties between DNA probes and analyte molecules.

Systemic lupus erythematosus after coronavirus disease-2019 (COVID-19) infection: Case-based review

BackgroundCoronavirus disease-2019 (COVID-19) is a novel infectious disease, which presents with various clinical manifestations. There is growing evidence of an association between COVID-19 infection and autoimmune diseases. The aim of this case report was to demonstrate the association of COVID-19 infection and the development of systemic lupus erythematosus (SLE). Case presentationA 38 year old Iranian woman presented with progressive icterus, pleuritic chest pain, palpitation, dyspnea, photosensitivity and arthralgia 18-days after COVID-19 symptoms proved by a positive polymerized chain reaction (PCR). The chest and abdomen computerized tomography (CT) scan showed pericardial and pleural effusion and enlarged liver and abdominal lymph nodes. Antinuclear antibody (ANA), anti-double stranded deoxyribonucleic acid (anti-ds DNA) antibody and perinuclear anti-neutrophil cytoplasmic antibody (P-ANCA) were positive. She was diagnosed as SLE and was successfully treated with prednisolone 30 mg daily, hydroxychloroquine 200 mg daily and azathioprine 150 mg daily and she remarkably improved. Repeated anti-ds DNA antibody was positive. Due to nausea and abdominal discomfort, azathioprine was discontinued and replaced with mycophenolate mofetil 1500 mg daily. In the article, similar cases were presented; the mean interval between COVID symptoms and SLE presentations was 24.86 days. Pulmonary and renal involvements were the most common presentations of SLE triggered by COVID-19. The most frequently reported autoantibody was ANA ConclusionIt is necessary to be aware of the development of lupus disease in COVID-19 infected patients, because prompt diagnosis and treatment is very important to improve their outcome.

Analysis of Indigenous Bacteria as Microplastic Degradation of Sediment in the Sea Waters of Dumai, Riau Province

Nowdays Marine pollution resulted by microplastics is a global concern. Microplastic particles have the potential to cause damage to biota. One of solution to overcome the problem of marine pollution by microplastics by applying indigenous bacteria as microplastic degradation. This study was conducted to determine the type and ability of indigenous bacteria in sediments that are able to degrade microplastics. The research was conducted from October to December 2020. Sediment sampling was conducted in Dumai Sea Waters and sample analysis was conducted at the Marine Microbiology Laboratory, Faculty of Fisheries and Marine, Riau University. Indigenous bacteria from sediments were isolated in bacterial breeding media and then tested degradation capabilities on PET plastic samples. The result of the degradation test obtained ISL8 was able to degrade the mycoplastic with a percent degradation about 7%. Molecular test of ISL8 using PCR (Polymer chain reaction) 16S rRNA method and GenBank analysis is known that ISL8 is Bacillus sp

Routine wastewater-based monitoring of antibiotic resistance in two Finnish hospitals: focus on carbapenem resistance genes and genes associated with bacteria causing hospital-acquired infections

Wastewater-based monitoring represents a useful tool for antibiotic resistance surveillance. To investigate the prevalence and abundance of antibiotic resistance genes (ARGs) in hospital wastewater over time. Wastewater from two hospitals in Finland (HUS1 and HUS2) was monitored weekly for nine weeks (weeks 25-33) in summer 2020. A high-throughput real-time polymerization chain reaction (HT-qPCR) system was used to detect and quantify 216 ARGs and genes associated with mobile genetic elements (MGEs), integrons, and bacteria causing hospital-acquired infections (HAIs), as well as the 16S rRNA gene. Data from HT-qPCR were analysed and visualized using a novel digital platform, ResistApp. Eight carbapenem resistance genes (blaGES, blaKPC, blaVIM, blaNDM, blaCMY, blaMOX, blaOXA48, and blaOXA51) and three genes associated with bacteria causing HAIs (Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa) were studied. There was a significantly higher number of ARGs at both hospitals in weeks 27-30 (174-191 genes) compared to other sampling weeks (151-171 genes). Our analyses also indicated that the two hospitals, which used different amounts of antibiotics, had significantly different resistance gene profiles. Carbapenem resistance genes were more prevalent and abundant in HUS1 than HUS2. Across both hospitals, blaGES and blaVIM were the most prevalent and abundant. There was also a strong positive association between blaKPC and K.pneumoniae in HUS1 wastewater. Routine wastewater-based monitoring using ResistApp can provide valuable information on the prevalence and abundance of ARGs in hospitals. This helps hospitals understand the spread of antibiotic resistance in hospitals and identify potential areas for intervention.

COVID-19 in patients with Behçet's disease: Outcomes and rate of Behçet's exacerbations in a retrospective cohort.

ABSTRACTObjectivesAim of this study is to investigate the outcomes of coronavirus disease 2019 (COVID-19) in our cohort of Behçet’s disease (BD) patients in means of mortality, admission to hospital and/or intensive care unit and length of hospital stay, additionally, to reveal rate of BD exacerbations due to COVID-19.MethodsA retrospective cohort was formed from patients who have previously been followed with a diagnosis of BD. Patients of this cohort were retrospectively evaluated for a positive severe acute respiratory syndrome – coronavirus 2 (SARS-CoV 2) polymerized chain reaction (PCR) test result. In PCR positive patients, demographics, comorbid diseases, active medical treatment for BD and information regarding hospitalization, intensive care unit admission, mortality were collected from medical records. PCR positive patients were reached via phone numbers and Behçet’s Disease Current Activity Form (BDCAF) scores for pre and post-COVID-19 BD symptoms were calculated.ResultsOut of a total 648 BD patients, 59 were detected to have a positive SARS-CoV 2 PCR test. 88.1% of these patients were under colchicine treatment and 28.8% of patients were under azathioprine treatment. 3 of the 59 patients (5.0%) were found to be hospitalized, none of them was admitted to the intensive care unit or died. An increasing trend in the frequency of comorbid diseases and older age was observed in hospitalized patients. 32.2% of BD patients suffered from exacerbation of at least one symptom related to BD when evaluated with BDCAF, arthralgia/arthritis being the most common.ConclusionsWe observed no intensive care unit admission or mortality with COVID-19 in our BD patient cohort. A substantial number of patients suffered from exacerbation of BD symptoms.

A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS.

Background:Concern has been raised about DNA extraction from Mycobacterium tuberculosis due to its complex procedure. This study demonstrates a simple and fast DNA extraction method of mycobacterial genome to subsequent molecular investigation, such as Polymerization Chain Reaction (PCR) amplification, with species-specific primers and sequencing.Materials and Methods:Total DNA was isolated from M. tuberculosis cultured by using boil method. DNA was evaluated via measures of DNA quantity and quality (absorbance at 230, 260 and 280 nm), DNA integrity (electrophoresis). Molecular tests were tested namely PCR and sequencing.Conclusions:The quality of DNA obtained is acceptable for PCR and sequencing analysis. These findings demonstrate that the method used is inexpensive and suitable for minimum infrastructure facilities.

A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS

Background: Concern has been raised about DNA extraction from Mycobacterium tuberculosis due to its complex procedure. This study demonstrates a simple and fast DNA extraction method of mycobacterial genome to subsequent molecular investigation, such as Polymerization Chain Reaction (PCR) amplification, with species-specific primers and sequencing. Materials and Methods: Total DNA was isolated from M. tuberculosis cultured by using boil method. DNA was evaluated via measures of DNA quantity and quality (absorbance at 230, 260 and 280 nm), DNA integrity (electrophoresis). Molecular tests were tested namely PCR and sequencing. Conclusions: The quality of DNA obtained is acceptable for PCR and sequencing analysis. These findings demonstrate that the method used is inexpensive and suitable for minimum infrastructure facilities.