Development Of Diagnostic Tests Research Articles

PROSPECTS AND CHALLENGES OF PHARMACEUTICAL BIOTECHNOLOGY

Biotechnology is a broad area of biology, involving the use of living systems and organisms to develop products. Depending on the tools and applications, it often overlaps with related scientific fields. In the late 20th and early 21st centuries, biotechnology has expanded to include new and diverse sciences, such as genomics, recombinant gene techniques, applied immunology, and development of pharmaceutical therapies and diagnostic tests. Biotechnology has also led to the development of antibiotics. Biotechnology has applications in four major industrial areas, including health care (medical), crop production and agriculture, non-food (industrial) uses of crops and other products and environmental uses. In medicine, modern biotechnology has many applications in areas such as pharmaceutical drug discoveries and production, pharmacogenomics, and genetic testing. Pharmaceutical biotechnology is a relatively new and growing field in which the principles of biotechnology are applied to the development of drugs. A majority of therapeutic drugs in the current market are bio formulations, such as antibodies, nucleic acid products and vaccines. Such bio formulations are developed through several stages that include: understanding the principles underlying health and disease the fundamental molecular mechanisms governing the function of related biomolecules synthesis and purification of the molecules determining the product shelf life, stability, toxicity and immunogenicity drug delivery systems patenting and clinical trials. This review article describes the purpose of biotechnology in pharmaceutical industry, particularly pharmaceutical biotechnology along with its prospects and challenges.

Flexible sensor based on conducting polymer and gold nanoparticles for electrochemical screening of HPV families in cervical specimens

Considering the low sensitivity of cytological exams and high costs of the molecular methods, the development of diagnostic tests for effective diagnosis of HPV infections is a priority. In this work, biosensor composed of polypyrrole (PPy) films and gold nanoparticles (AuNPs) was obtained for specific detection of HPV genotypes. The biosensor was developed by using flexible electrodes based on polyethylene terephthalate (PET) strips coated with indium tin oxide (ITO). Polymeric films and AuNPs were obtained by electrosynthesis. Oligonucleotides sequences modified with functional amino groups were designed to recognize HPV gene families strictly. The modified oligonucleotides were chemically immobilized on the nanostructured platform. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used for the analysis of the electrode modification and monitoring of molecular hybridization. Electrochemical changes were observed after exposure of the biosensors to plasmid samples and cervical specimens. The biosensor based on the BSH16 probe showed a linear concentration range for target HPV16 gene detection of 100 pg μL−1 to 1 fg μL−1. A limit of detection (LOD) of 0.89 pg μL−1 and limit of quantification (LOQ) of 2.70 pg μL−1 were obtained, with a regression coefficient of 0.98. Screening tests on cervical specimens were performed to evaluate the sensibility and specificity for HPV and its viral family. The expression of a biomarker for tumorigenesis (p53 gene) was also monitored. In this work, a flexible system has been successfully developed for label-free detection of HPV families and p53 gene monitoring with high specificity, selectivity, and sensitivity.

Efficacy and Safety of AZD1222, BNT162b2 and mRNA-1273 vaccines against SARS-CoV-2

SARS-CoV-2, the beta coronavirus causing COVID-19, was isolated and categorizes as a novel one on January 7th, 2020 in China.[1] To date, official reports depict that SARS-CoV-2 has already infected 88.828.387 persons and caused 1.926.625 deaths worldwide.[2] On January 12th, 2020, China officials made public its genetic sequence, thus paving the way towards the research and development of diagnostic tests and vaccines.
 With regard to vaccination, e large number of clinical trials were designed and are currently undergoing, of which 189 are listed in ClinicalTrials.gov. [3] However, up to date, only three vaccines have published their respective phase III clinical trial results in peer-reviewed medical journals. [4-6]
 Vaccines are needed to reduce the morbidity and mortality associated with Covid-19, and multiple vaccine platforms as AZD1222 (AstraZeneca) [4], BNT162b2 (Pfizer/BioNTech) [5] and mRNA-1273 (Moderna) have been involved in the rapid development of vaccine candidates.
 Methodology: In this review, PubMed, Embase, Web of Science, Scopus, medRxiv, and bioRxiv were systematically scrutinized for peer-reviewed and preprint articles on phase III clinical trials of vaccines against SARS-CoV-2. In total, only three peer-reviewed papers fulfilling the search criteria were identified.
 Conclusions; All vaccine candidates should publish in peer-reviewed journals their efficacy and safety well before requesting approval to the national or international authorities…

Basic principles of nuclear techniques and applications in livestock production

It is known. that many of the advances which have been made in improving the feeding, fertility and health of livestock in developed countries would not have been possible without nuclear tech-niques. Nuclear techniques have been used by scientists to study and understand those biological processes which are responsible for growth, reproduction and disease resistance in animals. Tracer tech-niques based on isotopes have been used for tagging feed constituents, amino acids and other metabolites, whereby their fate .can be fol-lowed within the digestive system and other parts of the body. This forms an integral part of the armoury used by animal nutritionists to establish the nutri-tional value of all the materials potentially available to feed animals. The development and use of highly sensi­tive immunoassay methods, to measure the minute quantities. of reproductive hormones in the blood enables us to determine when. animals are ready- for breeding, check whether animals have been inseminated at the correct time, diagnose pregnancy early enough, and devise corrective measures for reproductive disorders and improve the efficiency of artificial insemination and embryo transfer programmes. Radioisotopic methods have been used to study the immune system. to examine infec­tious agents and how they Stimulate this system, and to develop specific and sensitive methods for development of veterinary vaccines and diagnostic tests.

Handheld Point-of-Care System for Rapid Detection of SARS-CoV-2 Extracted RNA in under 20 min.

The COVID-19 pandemic is a global health emergency characterized by the high rate of transmission and ongoing increase of cases globally. Rapid point-of-care (PoC) diagnostics to detect the causative virus, SARS-CoV-2, are urgently needed to identify and isolate patients, contain its spread and guide clinical management. In this work, we report the development of a rapid PoC diagnostic test (<20 min) based on reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) and semiconductor technology for the detection of SARS-CoV-2 from extracted RNA samples. The developed LAMP assay was tested on a real-time benchtop instrument (RT-qLAMP) showing a lower limit of detection of 10 RNA copies per reaction. It was validated against extracted RNA from 183 clinical samples including 127 positive samples (screened by the CDC RT-qPCR assay). Results showed 91% sensitivity and 100% specificity when compared to RT-qPCR and average positive detection times of 15.45 ± 4.43 min. For validating the incorporation of the RT-LAMP assay onto our PoC platform (RT-eLAMP), a subset of samples was tested (n = 52), showing average detection times of 12.68 ± 2.56 min for positive samples (n = 34), demonstrating a comparable performance to a benchtop commercial instrument. Paired with a smartphone for results visualization and geolocalization, this portable diagnostic platform with secure cloud connectivity will enable real-time case identification and epidemiological surveillance.

Updates from the 2020 World Health Organization Classification of Soft Tissue and Bone Tumours.

The fifth edition of the World Health Organization (WHO) classification of soft tissue and bone tumours was published in May 2020. This 'Blue Book', which is also available digitally for the first time, incorporates an array of new information on these tumours, amassed in the 7years since the previous edition. Major advances in molecular characterisation have driven further refinements in classification and the development of ancillary diagnostic tests, and have improved our understanding of disease pathogenesis. Several new entities are also included. This review summarises the main changes introduced in the 2020 WHO classification for each subcategory of soft tissue and bone tumours.

A Highly Specific DNA Aptamer for RNase H2 from Clostridium difficile.

Molecular recognition elements with high specificity are of great importance for the study of molecular interactions, accurate diagnostics, drug design, and personalized medicine. Herein, a highly specific DNA aptamer for RNase H2 from Clostridium difficile (C. difficile) was generated by SELEX and minimized to 40 nucleotides. The aptamer exhibits a dissociation constant (Kd) of 1.8 ± 0.5 nM and an inhibition constant (IC50) of 7.1 ± 0.6 nM for C. difficile RNase H2, both of which are 2 orders of magnitude better for the same enzyme from other control bacteria. The fluorescent version of the aptamer can distinguish C. difficile from several other control bacteria in a cell lysate assay. This work demonstrates that a ubiquitous protein like RNase H2 can still be used as the target for the development of highly specific aptamers and the combination of the protein and the aptamer can achieve the recognition specificity needed for a diagnostic test and drug development.

Functional Comparison of Bioactive Cellulose Materials Incorporating Engineered Binding Proteins.

Whatman No. 1 chromatography paper is widely used as a substrate for cellulose-based immunoassays. The immobilized proteins are used to capture target biomarkers for detection. However, alternative paper substrates may facilitate mass production of immunoassays as diagnostic tests. Here, we assessed the physical characteristics and protein immobilization capabilities of different commercial papers. Some substrates fulfilled our design criteria, including adequate flow rate and sufficient protein immobilization for efficient target capture. This study demonstrates that a variety of paper substrates can be bioactivated and used to capture target biomarkers, enabling development of affordable diagnostic tests from a range of starting materials.

Non-inflammatory pancreatic cysts: from diagnosis to treatment (97 cases series).

to describe the implications of the diagnosis and treatment of non-inflammatory pancreatic cysts in a series of patients. we included patients with pancreatic cysts ≥1.0 cm, excluding those with a presumptive diagnosis of a pseudocyst. Imaging tests, echoendoscopy, and histopathology determined the diagnosis of the type of cyst. We applied the guidelines of the International Association of Pancreatology, with some modifications, in patients with mucinous or indeterminate lesions. 97 adult patients participated in the study. A cystic neoplasm of the pancreas was diagnosed in 82.5% of cases. Diagnosis was mainly made by magnetic resonance (46% of cases). The two most common diagnoses were intraductal papillary mucinous neoplasm (43.3%) and serous cystadenoma (26%). Twenty-nine patients underwent surgery (33.3%). The most common surgical procedure was distal pancreatectomy associated with splenectomy in 19 cases (65.5%). Among the operated patients, 11 were diagnosed with cancer. None of the followed, non-operated patients had a diagnosis of cancer. magnetic resonance showed good accuracy, particularly in the diagnosis of intraductal papillary mucinous neoplasm. The guidelines of the International Association of Pancreatology, as applied in this study, showed a negative predictive value for cancer of 100%. A development of better diagnostic tests can reduce the number of unnecessary operations.

The role of national and international veterinary laboratories.

In the field of diagnostic test validation, World Organisation for Animal Health (OIE) Reference Laboratories (RLs) have a pivotal role and provide the international community with impartial advice and support in the selection, development and validation of diagnostic tests, which can be applied to the specialist diseases for which they are designated. National RLs provide an invaluable function in supporting the introduction, ongoing validation and application of validated diagnostic tests in line with international standards. Experienced staff with extensive knowledge of such systems and access to specialist facilities for conducting work are available to monitor changes or advancements in technology. They consider their relevance and value to evolving diagnostic test requirements. Reference Laboratories often have a broad mandate of activity linking research or development programmes and surveillance activities to benefit the continual assessment and, if necessary, improvement of diagnostic tools. Reference Laboratories maintain or have access to unique biological archives (known positive and negative sample populations) and produce international reference standards, both of which are vital in establishing the necessary and detailed validation of any diagnostic test. Reference Laboratories act either singularly or in collaborative partnerships with other RLs or science institutes, but also, when required, and with impartiality, with the commercial sector, to ensure new tests are validated according to OIE standards. They promote and apply formal programmes of quality assurance (including proficiency testing programmes) for newly validated tests, ensuring ongoing monitoring and compliance with standards, or as required set out any limitations or uncertainties. Reference Laboratories publish information on test validation in the scientific literature and on relevant websites, as well as disseminating information at workshops and international conferences. Furthermore, they can offer training in the processes and systems underpinning test validation.

Validation, certification and registration of veterinary diagnostic test kits by the World Organisation for Animal Health Secretariat for Registration of Diagnostic Kits.

In the field of diagnostic test validation, World Organisation for Animal Health (OIE) Reference Laboratories (RLs) have a pivotal role and provide the international community with impartial advice and support in the selection, development and validation of diagnostic tests, which can be applied to the specialist diseases for which they are designated. National RLs provide an invaluable function in supporting the introduction, ongoing validation and application of validated diagnostic tests in line with international standards. Experienced staff with extensive knowledge of such systems and access to specialist facilities for conducting work are available to monitor changes or advancements in technology. They consider their relevance and value to evolving diagnostic test requirements. Reference Laboratories often have a broad mandate of activity linking research or development programmes and surveillance activities to benefit the continual assessment and, if necessary, improvement of diagnostic tools. Reference Laboratories maintain or have access to unique biological archives (known positive and negative sample populations) and produce international reference standards, both of which are vital in establishing the necessary and detailed validation of any diagnostic test. Reference Laboratories act either singularly or in collaborative partnerships with other RLs or science institutes, but also, when required, and with impartiality, with the commercial sector, to ensure new tests are validated according to OIE standards. They promote and apply formal programmes of quality assurance (including proficiency testing programmes) for newly validated tests, ensuring ongoing monitoring and compliance with standards, or as required set out any limitations or uncertainties. Reference Laboratories publish information on test validation in the scientific literature and on relevant websites, as well as disseminating information at workshops and international conferences. Furthermore, they can offer training in the processes and systems underpinning test validation.

Deep learning for COVID-19 chest CT (computed tomography) image analysis: A lesson from lung cancer

As a recent global health emergency, the quick and reliable diagnosis of COVID-19 is urgently needed. Thus, many artificial intelligence (AI)-base methods are proposed for COVID-19 chest CT (computed tomography) image analysis. However, there are very limited COVID-19 chest CT images publicly available to evaluate those deep neural networks. On the other hand, a huge amount of CT images from lung cancer are publicly available. To build a reliable deep learning model trained and tested with a larger scale dataset, the proposed model builds a public COVID-19 CT dataset, containing 1186 CT images synthesized from lung cancer CT images using CycleGAN. Additionally, various deep learning models are tested with synthesized or real chest CT images for COVID-19 and Non-COVID-19 classification. In comparison, all models achieve excellent results in accuracy, precision, recall and F1 score for both synthesized and real COVID-19 CT images, demonstrating the reliable of the synthesized dataset. The public dataset and deep learning models can facilitate the development of accurate and efficient diagnostic testing for COVID-19.

Modifying laboratory testing via home brew during the COVID-19 pandemic.

Rapid development and deployment of diagnostic testing for COVID-19 have been a key component of the public health response to the pandemic. Out of necessity, academic and other clinical laboratories developed laboratory testing innovations for COVID-19 to meet clinical testing demands. In addition to constraints on local testing supplies and equipment, a rapidly changing regulatory framework created challenges for translational scientists. Illustrative examples of approaches used to develop laboratory tests during the early stages of the COVID-19 pandemic demonstrate effective team science approaches to this challenging clinical care and public health emergency. These experiences and the associated lessons learned are relevant to the development of public health response plans for future pandemics.

Nanobiosensors for Pathogenic Agents Detection

Conventional methods to detect pathogenic microorganisms can be wasteful, expensive, and not very sensitive. Due to these limitations, approaches based on nanotechnology and the development of new diagnostic tests are emerging as alternative methods. Resulting from the unique electrical, magnetic, luminescent, and catalytic properties of nanomaterials, rapid and cost-effective diagnostic tests have been developed to recognize the presence of microbial pathogens or some of their components in different samples. The purpose of this review is to provide an overview and an update regarding the use of several nanomaterials as biosensors, among which are gold nanoparticles, carbon nanotubes, magnetic nanoparticles, and quantum dots. The application of these biosensors has opened a new interdisciplinary frontier between biological detection and materials science.

The Importance of Diagnostic and Prognostic Biomarker Identification and Classification Towards Understanding ALS Pathogenesis.

ALS is increasingly perceived as a multisystem neurodegenerative disorder, and the identification of a panel of biomarkers that accurately reflect features of pathology is a priority, not only for diagnostic purposes but also for prognostic or predictive applications [1]. Thus, as a multisystemic disease, it is likely that a panel of biomarkers will be needed to fully capture the features of ALS pathology. Taking also into consideration the fact that its causes remain unknown to their majority, it remains a complex disease driven by a combination of several systemic parameters [2]. Beyond the monogenic causes, representing the 15% of the ALS cases, which list up to 30 gene mutations with the most frequent being C9orf72, SOD1, FUS, and TARDBP/TDP43 [3-5], much research is being done to identify and associate possible causes for the 80% of ALS cases (sALS and fALS combined) which at the moment are not explained by a known mutation [2, 4]. ALS sporadic cases are related to a multigenic component, and/or involve epigenetic modification, and/or result from DNA damage, environmental risk factors, behavioural factors, oxidative stress or viral infections leading to cellular dysfunctions [4, 6-10]. ALS diagnosis is lengthy and there is a typical diagnostic delay of 9-15months from onset to diagnostic confirmation based on clinical and electrophysiological criteria as well as the exclusion of diseases with similar behaviour to ALS. Three major exclusion criteria are involved in the diagnosis process: the Revised El Escorial Criteria (REEC), the Airlie House Criteria (AHC) and the Awaji Criteria [11, 12]. Taking into consideration that the average survival is 2-4years, it makes it urgent for the researchers to improve diagnostic speed and accuracy for ALS [13, 14]. In the absence of a reliable diagnostic test for ALS, biomarkers are a strong weapon not only for its diagnosis but also for understanding the pathomechanism as well as a basis for the development of therapeutics. Recent global research has accepted the fact that biomarkers will facilitate the combination of therapeutics with diagnostics and will thus play an important role in the development of personalized medicine [15]. This paper proposes a combination of diagnostic and prognostic biomarkers to meet the scope of global research. Thus, biomarkers specific to ALS pathology need to be identified towards the development of a reliable fast diagnostic test, while at the same time prognostic biomarkers should also be identified to monitor the status of the pathology as various candidates may serve both purposes. Finally, since different sub-cohorts of ALS patients respond differently to treatments [16], the identification of ALS biomarkers will contribute to a better understanding of the disease pathogenesis and permit targeted drug development and patient stratification leading to more efficient clinical trials.

Ketosis and its role in Bos taurus reproductive impairment

The analysis of relevant and significant scientific research on the physiological and biochemical aspects of ketosis, and the main mechanisms of influence on reproductive function in this metabolic pathology is presented. Strengthening breeding potential and creating conditions for its implementation through modern feeding and maintenance technologies led to the spread of diseases of non-communicable etiology, including ketosis diseases. Ketotic diseases cause various disorders in carbohydrate-lipid, protein, water-electrolyte and vitamin-mineral metabolism. The monitoring of studies on the biological role of ketosis showed its effect on animal fertility, quality of female gametes and fetal development. When considering ketosis, it is important to understand precisely the mechanisms of influence, since many methods for diagnosing and combating ketosis diseases are based on their knowledge. The key mechanisms of ketone bodies formation, the reasons for increase in their concentration were considered. It is of great practical importance for development of diagnostic tests that make it possible to determine the direction of disturbances in energy and plastic processes. Based on the data presented in the studies, conclusions were drawn about the role of -hydroxybutyric acid as a direct factor in reducing the reproductive function of Bos taurus .

Sequencing Analysis and Phylogenetic Tree of HPV Isolated from Breast Cancer Patients at Thi-Qar Province/Iraq

Genetic variety examination has demonstrated fundamental to the understanding of the epidemiological and developmental history of Papillomavirus (HPV), for the development of accurate diagnostic tests and for efficient vaccine design. The HPV nucleotide diversity has been investigated widely among high-risk HPV types. To make the nucleotide sequence of HPV and do the virus database in Thi-Qar province, and compare sequences of our isolates with previously described isolates from around the world and then draw its phylogenetic tree, this study done. A total of 6 breast formalin-fixed paraffin-embedded (FFPE) of the female patients were included in the study, divided as 4 FFPE malignant tumor and 2 FFPE of benign tumor. The PCR technique was implemented to detect the presence of HPV in breast tissue, and the real-time PCR used to determinant HPV genotypes, then determined a complete nucleotide sequence of HPV of L1 capsid gene, and draw its phylogenetic tree. The nucleotide sequencing finding detects a number of substitution mutation (SNPs) in (L1) gene, which have not been designated before, were identified once in this study population, and revealed that the HPV16 strains have the evolutionary relationship with the South African race, while, the HPV33 and HPV6 showing the evolutionary association with the North American and East Asian race, respectively.

Sequencing Analysis and Phylogenetic Tree of HPV Isolated from Breast Cancer Patients at Thi-Qar Province/Iraq

Genetic variety examination has demonstrated fundamental to the understanding of the epidemiological and developmental history of Papillomavirus (HPV), for the development of accurate diagnostic tests and for efficient vaccine design. The HPV nucleotide diversity has been investigated widely among high-risk HPV types. To make the nucleotide sequence of HPV and do the virus database in Thi-Qar province, and compare sequences of our isolates with previously described isolates from around the world and then draw its phylogenetic tree, this study done. A total of 6 breast formalin-fixed paraffin-embedded (FFPE) of the female patients were included in the study, divided as 4 FFPE malignant tumor and 2 FFPE of benign tumor. The PCR technique was implemented to detect the presence of HPV in breast tissue, and the real-time PCR used to determinant HPV genotypes, then determined a complete nucleotide sequence of HPV of L1 capsid gene, and draw its phylogenetic tree. The nucleotide sequencing finding detects a number of substitution mutation (SNPs) in (L1) gene, which have not been designated before, were identified once in this study population, and revealed that the HPV16 strains have the evolutionary relationship with the South African race, while, the HPV33 and HPV6 showing the evolutionary association with the North American and East Asian race, respectively.

COVID-19: concern about interrupting social isolation of healthcare workers and professionals. What should be done with the results of the available COVID-19 diagnostic tests?

Management of SARS-CoV-2 requires safe decision-making to minimize contamination. Healthcare workers and professionals in confined areas are affected by the risk of the activity and the environment. Isolation of contaminated workers and healthcare professionals requires clinical and diagnostic criteria. On the other hand, interrupting the isolation of healthcare employees and professionals is critical because diagnostic tests do not support clinical decisions. In addition to defining the best test in view of its accuracy, it is necessary to consider aspects such as the stage of the disease or cure, the viral load and the individual's own immunity. Uncertainty about natural and herd immunity to the disease leads to the development of appropriate antivirals, diagnostic tests and vaccines.

Development of a Four-Tier Multiple-Choice Test on The Concept of Transport Across Membranes

The development and use of diagnostic tests in the form of multiple choice four levels in biology learning are still lacking. This study aims to develop a valid and reliable four-tier multiple-choice test instrument to identify the conception profile of high school students on the concept of transport across membranes. The research method is in the form of development research by adapting the Kilic and Saglam models which consist of three stages, namely defining content, obtaining information on student misunderstandings, and developing tests. The research instruments used were validation sheets, test instruments, and interview sheets. Descriptive statistical analysis techniques were used to analyze the data in this study. The results showed that 15 items developed were declared valid and the reliability coefficient of the test was 0.82, including the very reliable category. Thus, it can be concluded that the four-tier multiple-choice test instrument developed has met the validity and reliability requirements to identify the conception profile of high school students about the concept of transport across membranes. The four-tier multiple-choice test developed can be used by high school teachers as an accurate test to identify the conception profile of their students because it can distinguish between students who understand the concept, do not understand the concept, or who experience misconceptions about the concept of transport across membranes.