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During this current coronavirus disease 2019 (COVID-19) pandemic, RT-PCR plays an essential role as a gold standard for the detection of SARS-CoV-2. This standard RT-PCR procedure, particularly the RNA extraction/ purification step, is a laborious and time-consuming workflow. Herein, we examined the extraction-free RT-PCR method combined with preheated step (95 °C for 5 min) to accelerate the process. From our result, direct RT-PCR method (with/without heating step) achieved 100 % specificity. Importantly, the additional heating step could remarkably improve the test's sensitivity (84 %) compared to direct RT-PCR without any preheated stage (66 %), with negative detection cut off CT value of 30. The significant agreeability of this simple preheated RT-PCR method with the standard extraction-based RT-PCR procedure highlights its potency as an alternative testing method for SARS-CoV-2 detection. Thus, it may improve the testing capacity of COVID-19 screening and serve as a contingency for resource-challenged settings when extraction supplies are scarce. [ABSTRACT FROM AUTHOR]

COVID-19 pandemic originated in December 2019 due to SARS-CoV-2 virus infection. Standardized SARS-CoV-2 detection is examined by Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) with N and RdRp genes as the main target. Local SARS-CoV-2 detection product is not readily available with only N2, RdRp, and Orf1b as gene targets. This study aims to optimize multiplex RT-qPCR with two pairs of primer set targeting N and RdRp genes with RPP30 gene as internal control. The study was done using several methods: Escherichia coli culture production and plasmid extraction, HepG2 cell RNA extraction, PCR, RT-PCR, RT-qPCR, and electrophoresis. The result of the earlier steps was successful plasmid and RNA extraction. PCR on plasmid targeting N and RdRp genes and RT-PCR on RNA targeting RPP30 gene giving results of bands appearance with the size around 100 bp after electrophoresis is done. RT-qPCR of two pairs of primer set generally giving positive results with the appearance of logarithmic graph on RT-qPCR plots. The conclusion of the study is RT-qPCR was done successfully using two sets of primer that targeting N and RdRp genes with RPP30 gene as internal control. [ABSTRACT FROM AUTHOR]

Chrysanthemum (Dendranthema grandiflora) is a cut flower plant that has become a world commodity. The bright color is not only liked by the public but also attracts insects, including Aphis spp which is a vector of Chrysanthemum stunt viroid (CSVd) disease. As a result, the production and quality of the affected chrysanthemum are low. The most promising control method for the virus is planting CSVd-free chrysanthemum seeds. Therefore, it is necessary to explore the optimal molecular-based CSVd detection method for screening viroid-free chrysanthemum seedlings. Current approaches to detect CSVd return gel electrophoresis, dot-blot hybridization assay, and reverse transcription-polymerase chain reaction (RT-PCR). Of the eleven types of indicator plants tested, there were three types of plants that showed positive reactions to CSVd (based on RT-PCR), namely Senecio. v cruentus, Petunia hybrid and Dendranthema grandiflora F1 hybrid. Likewise with the return-PAGE technique as well as with the hybridization technique. The success of this detection was evidenced by the presence of purple spots in the area where the stem pieces of sick chrysanthemum were attached to the nylon membrane, and the signal was not found in healthy plants. Thus the presence of CSVd RNA in chrysanthemum plants can be detected using RT-PCR techniques, tissue blot hybridization, and return PAGE. Tissue blot hybridization is the most feasible technique in the chrysanthemum certification program because it can be applied to a large number of samples at the same time. [ABSTRACT FROM AUTHOR]

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that cause coronavirus disease-2019 (COVID-19), is an acute respiratory disease that causes a major impact on the world. Indonesia is one of the countries that has a high mortality rate of COVID-19. Based on the WHO and the CDC, molecular test method is the recommendation for diagnosing COVID-19. Real-time PCR test is an examination that is currently the gold standard for detecting SARS-CoV-2. There are currently many kits and reagents available to detect COVID-19, one of them is BioCoV-19 RT-PCR Kit from Biofarma that produced in Indonesia. The aim of this study was to compare basic analytical and clinical performance of BioCoV-19 RT-PCR kit with N1N2 CDC RT-PCR as the standard. In this study we used serial dilutions of SARS-CoV-2 viral RNA to establish PCR efficiency and estimate the limit of detection (LOD). Furthermore, we ran a panel of 135 clinical samples and compare the result for sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). Finally, we used clinical samples positive for non-coronavirus respiratory viral infections for cross reactions test. PCR efficiency of BioCoV-19 RT-PCR kit was ≥96 % and the estimated LOD within a 5-fold range. The assay does not showed cross-reactivity with other respiratory viruses. We conclude that real-time BioCoV-19 RT-PCR kit assessed in this study may be used for routine diagnostics of COVID-19 in patients. [ABSTRACT FROM AUTHOR]

It was discovered that Coronavirus disease of 2019 (COVID-19) caused the death of a multitude of lifes and caused health problems and weakness. The WHO has declared this a pandemic. Hundreds of thousands of people are infected every day with this virus. In order to aid in automating the scanning process, researchers are using medical images such as X-rays and computed tomography (CT) scans, and also AI-based systems, to assist in the diagnosis of COVID-19 due to the complexity and expense of conventional reverse transcription polymerase chain reaction (RT-PCR) methods. Here, we reviewed a number of those emerging AI-based models with the ability to recognize COVID-19 based on X-rays or CT scans of lungs. As of June 20, 2021, we examined research papers and collected information about available resources. In order to discover future research directions towards self-learning diagnosis of COVID diseases, also explored different datasets and analyzed experimental results of various algorithms. Researchers at the novice and beginner levels can use this survey to begin their work on COVID classification. [ABSTRACT FROM AUTHOR]

The COVID-19 (Corona virus disease 2019) outbreak induced millions of human beings the loss of life, still spread of the disease was unstoppable and was declared a pandemic with the aid of the WHO (World Health Organization). Therefore, human beings across the world are nevertheless getting contaminated every day. Reverse Transcription Polymerase Chain Reaction (RT-PCR) to identify COVID-19 is not feasible in terms of both cost and time of identification, which might cost the patient's life. Therefore, to make identification economical and feasible, researchers were attempting to use clinical images (x-ray and CT etc.,) to detect COVID-19 with the assist of Machine Learning (ML) and Artificial Intelligence (AI) approaches to aid in automating the identification of the pandemic. This paper helps in understanding some ML and AI approaches for detecting COVID-19 from clinical lung images. The amassed records about accessible research sources and inspected a complete of 30 research papers until august 2021. This paper includes the exploration and analysis of data sets, pre-processing methods, segmentation approaches, feature extraction, classification, and test effects, which can be useful for discovering future lookup instructions in the area of automated analysis of COVID-19 sickness the usage of AI-based frameworks. [ABSTRACT FROM AUTHOR]

In December 2019, an outbreak of a disease known as severe acute respiratory syndrome coronavirus 2, SARS-CoV-2 (COVID-19) was discovered in Wuhan, China that has caused the deaths of many people around the world. Early diagnosis is needed to control the geological spread of the COVID-19 outbreak. The standard protocol of real time reverse transcription polymerase chain reaction (RT-PCR) and enzyme immunosorbent assay (ELISA) were performed for COVID-19 detection. However, they are limited by tedious sample collection and preparation procedures, longer waiting times for test results and the lack of competent personnel to operate sophisticated lab-based instruments. In this context, the point-of-care (POC) diagnostic platform has proven to be a prospective approach in addressing the above challenges. Therefore, there is a growing urgency to develop diagnostic tools for rapid detection of coronavirus to prevent subsequent spread. This device is necessary and should be sensitive, selective, and demonstrates rapid diagnostic capabilities. Electrochemical DNA biosensors have exhibited these properties and, thus, serve as a promising candidate for virus detection. This review focuses on COVID-19 viral infection and physiopathology in depth. The electrochemical mechanism of the biosensor is then discussed, as well as its capacity to detect COVID-19 in this investigation. Finally, a focus on combining nanotechnology with smartphones to produce smart nano-biosensors was highlighted, which can promote COVID-19 diagnosis that is accessible to both people and the public in patients. [ABSTRACT FROM AUTHOR]

The many biologic effects of CCL5 gene are mediated by their interaction with chemokine receptors on the cell surface, which have of numerous physiological effects. Therefore, CCL5 may be linked to asthma exacerbation and may be associated as a chemokine related to a super family of secreted of a many proteins assessed to a tree of super family in role with immune-regulatory and infiltrating of inflamed cells. The study included 50 subjects, as in (40) patients taken from a numbered of (11) silos which workers that are exposed to grain dust and pesticide fumigants, which involved allergic patients, their ages ranged between (17-76) years. Which were suffered from allergic symptoms and sthma, vary between sever and modurate symptoms compared to (10) apparently healthy (18-72) years. Samples were obtained from workers whom operated in a central units of silos, that responsible for collection of grains in stores and evaporated by pesticide in a number of (11) silos. The samples tested for the levels of CCL5 gene expression by Quantitative expression of ccl5 gene and was determined by reverse transcription quantitative polymerase chain reaction (RT-qPCR), in which the relative quantitation method was employed. The results showed high significant elevation (p≤0.01) in the expression level of gene) in the patient that have allergic in comparison with healthy individual. The aim of the study to show the effect of CCL5 gene in allergic and asthma diseases specially in workers. [ABSTRACT FROM AUTHOR]

An outbreak of a unique coronavirus disease (i.e., COVID-19) has been recorded in Wuhan, China considering the fact that overdue December 2019, which ultimately has become a deadly disease across the world. Although COVID-19 is an acutely dealt with disease, it may additionally be deadly with a hazard of fatality of 4.03% in China and the best of 13.04Algeria and 12.67% in Italy (as of eighth April 2020). The onset of significant contamination may also bring about dying on account of vast alveolar harm and innovative breathing failure. Although laboratory trying out, e.g., the usage of reverse transcription polymerase chain reaction (RT-PCR), is the golden widespread for scientific prognosis, the checks may also produce fake negatives. Moreover, below the pandemic situation, a scarcity of RTPCR trying outsources may additionally put off the subsequent scientific choice and treatment. Under such circumstances, there are many methods for prognosis and analysis of COVID-19 patients. In this survey paper, various deep learning techniques for detection of COVID-19 are discussed. [ABSTRACT FROM AUTHOR]

Many people express their opinions regarding policies or rules imposed by the government through Twitter social media. This study was conducted to determine public sentiment regarding the rules for implementing the reverse-transcriptase polymerase chain reaction (RT-PCR) test as one of the requirements for using air transportation. The research was conducted using the Random Forest and K-Nearest Neighbor (KNN) algorithms and the CRISP-DM research method. Twitter data used is classified into negative, neutral and positive. The results of the sentiment analysis showed that the KNN algorithm outperformed Random Forest with an accuracy value of 75.63% and the sampling technique used was stratified sampling. The results of the sentiment analysis show that the public has a negative sentiment towards the policies issued by the government. [ABSTRACT FROM AUTHOR]

Detection tests for COVID-19 are important to mitigate its spread. However, it is not yet clear what are the best practices for its realization. This paper aims to propose best practices to avoid possible failures in the points of contact of the samples collection for Reverse Transcription Polymerase Chain Reaction (RT-PCR) process in an emergency department of a hospital for the care of patients who are suspected or diagnosed with the new coronavirus. For this purpose, we performed a process mapping of the RT-PCR collection process using the Service Blueprint tool to identify the points of contact between the patients and the hospital staff. Thus, we identified the possible failures in these points, with the process actors, and we prioritized them according to their perception about Gravity, Urgency, and Tendency matrix. Our results show that the most critical failures are related to the information and communication between the process actors and the patients. In this way, improvements were proposed related to the standardization, information material, continued training, development of specialized teams, creation of protocols, and use of information technology. [ABSTRACT FROM AUTHOR]

COVID 19 disease is causing a pandemic worldwide. As of now, reverse transcription-polymerase chain reaction (RT-PCR) test is the de facto test used to examine the existence of the ribonucleic acid (RNA) of SARS-CoV-27. RTPCR is one of the viable approaches to test for the disease. It is being used to test people for COVID 19 in most countries. Despite high specificity of testing with RT-PCR, this method has very low sensitivity. There is also high variability in efficacy as different methods of sampling and time of symptom occurence differ from person to person. The testing technique proposed as an essential tool for COVID-19 screening in this paper is especially effective as a complementary method to RT-PCR. COVID 19 infects mainly the lungs and alveoli. Lung CT scan images are being used. Therefore, it is essential for us to develop a machine learning code that can analyze lung computerized tomography images to detect presence of COVID 19. This is what we are trying to achieve by this project. [ABSTRACT FROM AUTHOR]

The recent outbreak of coronavirus (COVID-19) pandemic has exposed the weakness of the existing healthcare facilities in developing countries like Bangladesh. The increasing amount of patients has made this condition more vulnerable. There is a high possibility that, these increasing amount of symptomatic patients might create a shortage in RT-PCR test kits in upcoming days. The objective of this study is to use Define, Measure, Analyze, Improve, and Control (DMAIC) in improving the epidemic healthcare management system during the COVID-19 outbreak in Bangladesh. The goal of this study is to use LSS methodology, especially the DMAIC improvement format in the existing dedicated healthcare management system for coronavirus treatment. The root cause analysis behind the higher response time and improper service for the Institute of Epidemiology, Disease Control and Research (IEDCR), and other dedicated healthcare providers regarding, coronavirus treatment in the current situation has been performed. FMEA (Failure Mode and Effect Analysis) was conducted in order to assess the potential failure modes in the existing healthcare management system. A simulation study regarding the implementation of pooled testing in Bangladesh for improving efficiency and optimizing the usage of RT-PCR test kits has also been carried out. Subsequently, the feasibility study of implementing the pooled testing methodology was performed. Finally, recommendations and conclusions have been presented. [ABSTRACT FROM AUTHOR]

Cyclophosphamide is a common anticancer drug that belongs to the nitrogen mustard group of alkylating agents, their acts by adding an alkyl group to DNA. Due to this cytotoxic property it is used extensively in a variety of carcinomas singly or in combination with other drugs to treat a variety of cancers. Purinergic nucleotides targets a group of receptors known as purinergic receptors, which fall into two classes P2Y and P2X receptors, the P2Y receptors are membrane bound G protein coupled receptors. Purinergic signaling has been shown to play a significant role in the regulation of proliferation in many animal and human tissues. Therefore, the present study was conducted to compare and correlate histomorphological alterations in different anatomical sites of adult wistar albino rat induced by a gradual increase in cyclophosphoamide dose administered intraperitoneally and subcutaneously and correlate the effect of cyclophosphamide with purinergic receptor gene expression. The aim of this study is to examine the effect of cyclophosphoamide on histological and cellular changes when administered in two different methods as well as examine purinergic receptor gene expression post cyclophosphoamide injection. Material and methods: Laboratory wistar albino rats were injected with different bolus doses of cyclophosphoamide intraperitoneally and subcutaneously and were sacrificed after seven days. liver, kidney, testes and subcutaneous injection sites were examined histologically and compared to control animals. Purinergic receptors P2Y1, P2Y2 and P2Y4 Gene expression changes were assessed using Quantitative reverse transcriptase polymerase chain reaction. Results: histological examination of all tissues in both groups shows gradual cytotoxic effect of cyclophosphoamide reaching extreme histological damage with increased bolus dose. Tissue damage included hemorrhaging and lymphocytic collection. Gene fold change of purinergic receptors showed an increase in genetic expression of P2Y1, P2Y2 and P2Y4 receptors in all examined site which indicates ectopic overexpression of purinergic receptors with increases cyclophosphamide specific dose. Increased gene fold change is an indication to cellular response to inflammatory histological damage caused by cyclophosphamide administration. The results of this study show that purinergic receptors are highly involved in cellular damage restoration through ectopic gene overexpression in tissues damaged by cyclophosphamide. Conclusion: This study concludes that cyclophosphoamide has a strong cytotoxic effect on certain organs if injected at a high dose intraperitoneally of subcutaneously. However histological damage is more prominent in intraperitoneal injected animals compared to subcutaneously injected animals. Cytotoxic effect and histological damage is confirmed by the gradual increase in purinergic receptors gene expression in both intraperitoneally and subcutaneously injected animals indicating an inflammatory response by increased gene fold change post administration of cyclophosphoamide and with gradual increase in cyclophosphoamide dose. It is a safer and less damaging to administer cyclophosphoamide subcutaneously. [ABSTRACT FROM AUTHOR]

The research aims to detection the role of Epstein bar viruses (EBVs) that can be correlative with multiple sclerosis (MS) by identifying interactions between MS male patients by using immunological and molecular techniques, as well as the impact of various types of treatment lines (first line: Rebif, Avonex, Betaferon) and (second-line: Gilenya, Tysabri) on the subject of the study aspects. Enzyme-linked immunosuppression (ELISA) was used to assess serum levels in study groups; the results of this study revealed significantly increased antibody concentration of mean value of IL-17A in male patients than control respectively(105.96±16.30; 12.48±3.62 pg/ml), and EBV EBNA1IgG (0.05±0.02; 0.03±0.01 U/ml) in sera, while both melatonin hormone in MS male patient than control group respectively (0.59±0.12;1.10±0.13 pg/ml), and vitamin 03 (27.98±15.79; 41.90±7.22 ng/ml) show significant decrease than control groups which were in normal value, as well as the use of RT-PCR technique to estimate viral load from EBV(22.61±12.72 copies/ml). With respect to treatment lines, the results of this study showed that there was a significant difference between male patients and control groups for all parameters studied in this study, and by using ANOVA test it found the p-value for IL-17A was significantly different between control groups and both first and second-line treatment (p≤0.0001), where is no significant difference in other studied aspect. With respect to Expanded disability status score (E0SS), there is a significant difference between EDSS <3 and ≥3 for vitamin D3(p ≤0.0001).It could be concluded that EBV has a crucial role in the initiation and targeting MS disease, leading to increasing levels of IL-17A, and decrease serum levels of melatonin hormone for male patients. [ABSTRACT FROM AUTHOR]

SARS-CoV-2 is a highly contagious virus that can induce pulmonary complications like viral pneumonia and acute respiratory distress syndrome (ARDS). In order to support RT-PCR testing, chest X-rays are used to identify the presence of COVID-19 in lungs. Radiologists proficient in chest X-Ray (CXR) interpretation are scarce, motivating our work of examining the performance of convolutional neural networks (CNNs) on this task. CNNs are the state-ofthe- art image classification method. In this work we classify X-rays into four classes (COVID-19, other lung opacity, other diseases and normal). We find, that MobileNetV1 is the best CNN for this task and achieve an overall accuracy of 70% and a COVID-19 accuracy of 83% on a test data set. By increasing the number of images of the unrestricted classes normal, lung opacity and other, the COVID-19 accuracy can be increased to 95% and the overall accuracy to 78%. We leverage model interpretability techniques and provide attention heatmaps that can assist in validating the model's decision process. [ABSTRACT FROM AUTHOR]

In this century, global warming has become the main cause of temperature increase in various regions, including Indonesia. The increase of temperature gives negative effects to livestock such as the reduction of metabolism, reproduction, and performance. One of the most adaptive livestock to heat stress in Indonesia is Bali cattle. The response of Bali cattle to heat stress can be shown by the expression of HSP90 gene. This early study was conducted to understand the profile of HSP90 gene expression in Bali cattle to heat stress in West Sumbawa district, West Nusa Tenggara. Temperature humidity index (THI) was measured to ascertain the heat stress in Bali cattle that bred in West Sumbawa, West Nusa Tenggara and Cipelang district, Bogor as the control for five days in August 2016. The expression of HSP90 gene was observed from RNA isolation of blood samples of two years old Bali cattle, which were taken from venous jugularis. Transcript variants of HSP90 were analyzed by reverse transcription polymerase chain reaction (RT-qPCR). The results showed that THI measurement in West Sumbawa district is higher than in Cipelang district. Furthermore, there is a tendency that the ratio of HSP90 gene expression of Bali cattle in West Sumbawa district is higher than Cipelang district, as a control. Nevertheless, the values of the ratio of HSP90 gene expression in Bali cattle were not significantly different between the districts. However, this research showed that Bali cattle in West Sumbawa district, West Nusa Tenggara may have good adaptability to heat stress. [ABSTRACT FROM AUTHOR]

Recently, Euphorbiaceae have attracted much attention for their potential uses in biodiesel production. Their seeds have been well known as the principal resource for oil production. The seed-derived oil content ranged from 28 % to 39 % by dry weight for all Euphorbiaceae species. Meanwhile, their seed also possesses relatively high protein content, ranged from 22 % to 35 %, making them a new alternative for livestock animal feed resources. Nevertheless, the development of this new animal feed resource is limited to the toxicity of the seeds. Its toxicity is mainly caused by the presence of anti-nutrient agents and toxins, such as curcin, trypsin inhibitor and phorbol ester (PE). The later is known to be tetracyclic diterpenes, which represents the primary toxic substances in one of the Euphorbiaceae plants, Jatropha curcas L. To date, little is known about the biosynthesis of these toxic substances. This research aims to analyze the expression of JcGGPPS gene encoding the geranylgeranyl diphosphate synthase involved in the production of the primary precursor for diterpenoid substances. In addition, the phylogram was presented to show the Euphorbiaceae Casbene Synthase (CS). The RT-PCR analysis showed that JcGGPPS is expressed both in seeds and leaves of J. curcas. Meanwhile, in silico analysis of six Euphorbiaceae species showed that CS from Euphorbia esula, Euphorbia resinifera, Euphorbia peplus and Euphorbia fischeriana are highly identical. In addition, CS from J. curcas derives from a different origin than other Euphorbiaceae plants. [ABSTRACT FROM AUTHOR]

Salinity is one of the most severe environmental stresses affecting plant productivity worldwide. Studies of the physiological and molecular mechanisms involved in tolerance to salinity indicate that to prevent the accumulation and toxicity of sodium in leaf cytoplasm, plants have developed at least four mechanisms that function in a cooperative manner, i.e., restriction of Na+ influx, active Na+ extrusion at the root-soil interface, controlled translocation of Na+ to the shoots, and compartmentalization of Na+ into the vacuole. Ten barley genotypes showing contrasting phenotypes, selected from the USDA barley core collection (2750 lines) (accessions with high relative biomass under saline stress that have relatively high flag leaf Na+/K+ content, versus accessions with high relative biomass under saline stress that have low flag leaf Na+/K+ content) were cultivated in pots in hydroponic system under greenhouse conditions. Two salt stress treatments were applied to seedlings (7dsm-l and 15dsm-l) along with control (Odsm-1). All treatments were made in triplicate. The salinity was maintained for the rest of the growth cycle. Primers to amplify the barley candidate genes (HKT1;5, HKT1;4, NHX1, and V-H+-PPase) and to monitor their expression profile by semi-quantitative RT-PCR, were synthesized. This study will contribute to understanding the mechanism of salt tolerance in barley and provide alternative barley genotypes that are tolerant to salinity. [ABSTRACT FROM AUTHOR]

During COVID-19 Pandemic, RT-PCR is widely used for detection of SARS-CoV-2. 'RT' in RT-PCR has two definition, one is 'Reversed Transcription', and the other is 'Real-Time' which means amount of replicated genes corresponding timeline indicated as intensity of fluorescent light. The two definition represents main mechanism of RT-PCR. My research objective is figuring out which biological features of SARS-CoV-2 make it detectable by RT-PCR technique, and how RT-PCR works corresponding the features of SARS-CoV-2. My research helps clinical researchers understand clearly the mechanism of SARS-CoV-2 RT-PCR test, and it increases accuracy of examination procedure. Also, it helps understanding more about features of SARS-CoV-2. The data of SARS-CoV-2 from my research is useful in development of vaccine, or action plan towards the pandemic. RNA from COVID-19 patients' nasal swab samples was extracted. Master-mix (MMLV RTase, dNTPs, DNA polymerase, primer/probe sets for SARS-CoV-2 and phage control) was added to RNA. Subsequently, RT-PCR examination was conducted. After reversed transcription and spectrofluorimetry process in RT-PCR, light from fluorophores on probes of viral genes was detected. SARS-CoV-2 virus is retrovirus, doing reversed transcription from RNA to cDNA. It uses RNA dependent DNA polymerase. Instead of natural enzyme, MMLV was used in RT-PCR, and it worked successfully in reversed transcription of SARS-CoV-2. The first definition of 'RT', 'Reversed Transcription', matches the feature of retrovirus doing reversed transcription. Probe with quencher and fluorophore attaches to single strand DNA, and degrades by DNA polymerase. Near each other, quencher absorbs light from fluorophore. After degradation, quencher separates from fluorophore, and light is detected. In RT-PCR, cDNA replication was done successfully by polymerase, and light from fluorophore on probes of viral gene was detected. Therefore, 'Real Time' the second definition of 'RT' which means light detected corresponding timeline after replication, matches the feature of SARS-CoV-2 virus doing replication of cDNA. [ABSTRACT FROM AUTHOR]

This study examined the genetic basis and heredity of a proband that is a symptomatic female carrier of Duchenne Muscular Dystrophy with a deletion involving exon 45. The proband's family includes an unaffected father and older brother, while the mother and identical twin sister have the deletion but remain fully asymptomatic. As a recessive X-linked disease, the rarity of a symptomatic female for DMD cannot be understated. This study explored the method of inheritance in this family through various techniques. Creatine-Kinase levels were first examined in the proband and the twin sister to confirm that only one was truly symptomatic. X-inactivation testing led to a need to perform PCR of a large section of the X chromosome throughout the family to examine the possibility of recombination in the mother through further analysis of the amplified DNA. After determining the true mode of inheritance, RNA isolation from fibroblast cell lines, reverse transcriptase, and RT-PCR were performed to analyze expression levels in an attempt to discover the cause for the varied phenotypes between the twin daughters. With the findings of homologous recombination and highly skewed x-inactivation, this study gained further insight into a rare case with great implications for other x-linked illnesses and a further understanding of Duchenne Muscular Dystrophy. [ABSTRACT FROM AUTHOR]