scholarly journals Molecular and Immunological Diagnostic Techniques of Medical Viruses

2020 ◽  
Vol 2020 ◽  
pp. 1-19
Author(s):  
Daniel Hussien Reta ◽  
Tesfaye Sisay Tessema ◽  
Addis Simachew Ashenef ◽  
Adey Feleke Desta ◽  
Wajana Lako Labisso ◽  
...  
Keyword(s):  
Low Income ◽  
Viral Infections ◽  
Diagnostic Techniques ◽  
Diagnostic Methods ◽  
Low Income Countries ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
Molecular Diagnostic Techniques ◽  
Global Public Health ◽  
Patient Sample

Viral infections are causing serious problems in human population worldwide. The recent outbreak of coronavirus disease 2019 caused by SARS-CoV-2 is a perfect example how viral infection could pose a great threat to global public health and economic sectors. Therefore, the first step in combating viral pathogens is to get a timely and accurate diagnosis. Early and accurate detection of the viral presence in patient sample is crucial for appropriate treatment, control, and prevention of epidemics. Here, we summarize some of the molecular and immunological diagnostic approaches available for the detection of viral infections of humans. Molecular diagnostic techniques provide rapid viral detection in patient sample. They are also relatively inexpensive and highly sensitive and specific diagnostic methods. Immunological-based techniques have been extensively utilized for the detection and epidemiological studies of human viral infections. They can detect antiviral antibodies or viral antigens in clinical samples. There are several commercially available molecular and immunological diagnostic kits that facilitate the use of these methods in the majority of clinical laboratories worldwide. In developing countries including Ethiopia where most of viral infections are endemic, exposure to improved or new methods is highly limited as these methods are very costly to use and also require technical skills. Since researchers and clinicians in all corners of the globe are working hard, it is hoped that in the near future, they will develop good quality tests that can be accessible in low-income countries.

Diagnosis of melioidosis: the role of molecular techniques

2021 ◽  
Vol 16 (4) ◽  
pp. 271-288
Author(s):  
Ian Gassiep ◽  
Delaney Burnard ◽  
Michelle J Bauer ◽  
Robert E Norton ◽  
Patrick N Harris
Keyword(s):  
Laboratory Diagnosis ◽  
Diagnostic Techniques ◽  
Molecular Techniques ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
Molecular Diagnostic Techniques ◽  
Detection And Identification ◽  
Life Years ◽  
Culture Independent ◽  
Future Utility

Melioidosis is an emerging infectious disease with an estimated global burden of 4.64 million disability-adjusted life years per year. A major determinant related to poor disease outcomes is delay to diagnosis due to the fact that identification of the causative agent Burkholderia pseudomallei may be challenging. Over the last 25 years, advances in molecular diagnostic techniques have resulted in the potential for rapid and accurate organism detection and identification direct from clinical samples. While these methods are not yet routine in clinical practice, laboratory diagnosis of infectious diseases is transitioning to culture-independent techniques. This review article aims to evaluate molecular methods for melioidosis diagnosis direct from clinical samples and discuss current and future utility and limitations.

scholarly journals A Recent Update on Advanced Molecular Diagnostic Techniques for COVID-19 Pandemic: An Overview

2021 ◽  
Vol 12 ◽  
Author(s):  
Akanksha Roberts ◽  
Raghuraj Singh Chouhan ◽  
Deepshikha Shahdeo ◽  
Narlawar Sagar Shrikrishna ◽  
Veerbhan Kesarwani ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Electrochemical Sensors ◽  
Cost Effective ◽  
Surface Enhanced Raman Spectroscopy ◽  
Diagnostic Techniques ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Molecular Diagnostic Techniques ◽  
Rt Pcr ◽  
Ongoing Research

Coronavirus disease 2019 (COVID-19), which started out as an outbreak of pneumonia, has now turned into a pandemic due to its rapid transmission. Besides developing a vaccine, rapid, accurate, and cost-effective diagnosis is essential for monitoring and combating the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its related variants on time with precision and accuracy. Currently, the gold standard for detection of SARS-CoV-2 is Reverse Transcription Polymerase Chain Reaction (RT-PCR), but it lacks accuracy, is time-consuming and cumbersome, and fails to detect multi-variant forms of the virus. Herein, we have summarized conventional diagnostic methods such as Chest-CT (Computed Tomography), RT-PCR, Loop Mediated Isothermal Amplification (LAMP), Reverse Transcription-LAMP (RT-LAMP), as well new modern diagnostics such as CRISPR–Cas-based assays, Surface Enhanced Raman Spectroscopy (SERS), Lateral Flow Assays (LFA), Graphene-Field Effect Transistor (GraFET), electrochemical sensors, immunosensors, antisense oligonucleotides (ASOs)-based assays, and microarrays for SARS-CoV-2 detection. This review will also provide an insight into an ongoing research and the possibility of developing more economical tools to tackle the COVID-19 pandemic.

scholarly journals Prevalence of Bacterial Infections and the use of Multiplex Pcr Assay for Rapid Detection in Cultured Fish in Ghana.

2021 ◽  
Author(s):  
Rhoda Lims Diyie ◽  
Dennis W. Aheto ◽  
Mike Y. Osei-Atweneboana ◽  
Emmanuel Armah ◽  
Kobina Yankson
Keyword(s):  
Bacterial Infections ◽  
Molecular Genetic ◽  
Diagnostic Techniques ◽  
Infected Fish ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
Molecular Diagnostic Techniques ◽  
Stressed Condition ◽  
Multiplex Pcr Assay ◽  
Cultured Fish

Abstract The modern and rapid avenue for detecting pathogens provided by molecular genetic techniques including polymerase chain reaction (PCR) was explored in the present study to identify prevalent disease pathogens, from six aquaculture farms and in two commonly cultured fish in Ghana. The specific detection was carried out directly on clinical samples of naturally infected fish (O. niloticus and C. gariepinus) based on syber-mix reaction protocol in traditional PCR. Molecular diagnostic techniques allowed the detection of six most common and important bacteria pathogens in aquaculture farms in Ghana. Also, three of the pathogens (Streptococcus agalactiae, Streptococcus iniae and Staphylococcus aureus) were simultaneously isolated in a multiplex reaction. The results indicated 90% - 100% sensitivity and specificity for each of the six bacterial pathogens tested. Streptococcosis and motile aeromonad septicemia were found to be highly prevalent in most aquaculture farms in Ghana with severity in infections traced to the 85.7% and 14.9% co-infections with all six target pathogens in catfish and tilapia respectively. Prevalence rate of infections significantly correlated with variations in salinity, conductivity and dissolved oxygen concentrations in the thermal stressed condition of the culture water.

scholarly journals Human Parechovirus: an Increasingly Recognized Cause of Sepsis-Like Illness in Young Infants

2017 ◽  
Vol 31 (1) ◽  
Author(s):  
Laudi Olijve ◽  
Lance Jennings ◽  
Tony Walls
Keyword(s):  
Viral Infections ◽  
Severe Disease ◽  
Diagnostic Techniques ◽  
Molecular Diagnostic ◽  
Molecular Diagnostic Techniques ◽  
Human Parechovirus ◽  
Content Type ◽  
Cns Infections ◽  
Young Infants ◽  
Increased Risk

SUMMARYHuman parechovirus (HPeV) is increasingly being recognized as a potentially severe viral infection in neonates and young infants. HPeV belongs to the familyPicornaviridaeand is currently divided into 19 genotypes. HPeV-1 is the most prevalent genotype and most commonly causes gastrointestinal and respiratory disease. HPeV-3 is clinically the most important genotype due to its association with severe disease in younger infants, which may partly be explained by its distinct virological properties. In young infants, the typical clinical presentation includes fever, severe irritability, and rash, often leading to descriptions of “hot, red, angry babies.” Infants with severe central nervous system (CNS) infections are at an increased risk of long-term sequelae. Considering the importance of HPeV as a cause of severe viral infections in young infants, we recommend that molecular diagnostic techniques for early detection be included in the standard practice for the investigation of sepsis-like illnesses and CNS infections in this age group.

scholarly journals A Simple, Affordable, Rapid, Stabilized, Colorimetric, Versatile RT-LAMP assay to detect SARS-CoV-2

2020 ◽  
Author(s):  
Juan García-Bernalt Diego ◽  
Pedro Fernández-Soto ◽  
Marta Domínghez-Gil ◽  
Moncef Belhassen-García ◽  
Juan Luis Muñoz Bellido ◽  
...  
Keyword(s):  
Low Income ◽  
Lamp Assay ◽  
Low Income Countries ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
Technological Infrastructure ◽  
Testing Method ◽  
One Step ◽  
Alternative Testing ◽  
Temperature Storage

Abstract The SARS-Cov-2 pandemic has forced all countries worldwide to rapidly develop and implement widespread testing to control and manage the Coronavirus Disease 2019 (COVID-19). RT-qPCR is the gold standard molecular diagnostic method for COVID-19, mostly in automated testing platforms. These systems are accurate and effective, but also costly, time-consuming, high technological, infrastructure dependent and currently suffer from commercial reagent supply shortages. The reverse-transcription loop-mediated isothermal amplification (RT-LAMP) can be used as alternative testing method. Here, we present a novel versatile (real-time and colorimetric) RT-LAMP for the simple (one-step) and rapid (as soon as 9 min) detection of SARS-CoV-2 and demonstrate the assay on RT-qPCR-positive clinical samples. We further transformed the RT-LAMP into a dry format for room-temperature storage suitable for potentially ready-to-use COVID-19 diagnosis. After further testing and validation, the Dry-RT-LAMP could be easily applied both in developed and in low-income countries yielding rapid and reliable results.

scholarly journals A Simple, Affordable, Rapid, Stabilized, Colorimetric, Versatile RT-LAMP Assay to Detect SARS-CoV-2

Diagnostics ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 438
Author(s):  
Juan García-Bernalt Diego ◽  
Pedro Fernández-Soto ◽  
Marta Domínguez-Gil ◽  
Moncef Belhassen-García ◽  
Juan Luis Muñoz Bellido ◽  
...  
Keyword(s):  
Low Income ◽  
Reverse Transcription ◽  
Lamp Assay ◽  
Virus Genome ◽  
Low Income Countries ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
The Novel ◽  
Technological Infrastructure ◽  
One Step

The SARS-CoV-2 pandemic has forced all countries worldwide to rapidly develop and implement widespread testing to control and manage the Coronavirus Disease 2019 (COVID-19). reverse-transcription (RT)-qPCR is the gold standard molecular diagnostic method for COVID-19, mostly in automated testing platforms. These systems are accurate and effective, but also costly, time-consuming, high-technological, infrastructure-dependent, and currently suffer from commercial reagent supply shortages. The reverse-transcription loop-mediated isothermal amplification (RT-LAMP) can be used as an alternative testing method. Here, we present a novel versatile (real-time and colorimetric) RT-LAMP for the simple (one-step), affordable (~1.7 €/sample), and rapid detection of SARS-CoV-2 targeting both ORF1ab and N genes of the novel virus genome. We demonstrate the assay on RT-qPCR-positive clinical samples, obtaining most positive results under 25 min. In addition, a novel 30-min one-step drying protocol has been developed to stabilize the RT-LAMP reaction mixtures, allowing them to be stored at room temperature functionally for up to two months, as predicted by the Q10. This Dry-RT-LAMP methodology is suitable for potentially ready-to-use COVID-19 diagnosis. After further testing and validation, it could be easily applied both in developed and in low-income countries yielding rapid and reliable results.

scholarly journals Evaluation of designed IS711 primers and universal primers of B4 and B5 for detection of Brucella spp. in clinical samples

2020 ◽  
Author(s):  
Pedram Heidari ◽  
Mitra Salehi ◽  
Abbas Akhavan Sepahi ◽  
Mohamad Reza Razavi
Keyword(s):  
Diagnostic Techniques ◽  
The Other ◽  
Universal Primers ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
Molecular Diagnostic Techniques ◽  
Target Sequence ◽  
Molecular Approaches ◽  
Detection And Identification ◽  
Antibody Titers

Abstract Background: Brucellosis as a global concern is a zoonotic infectious disease which affects a large number of individuals in developing countries. Microbiological, serological and molecular approaches are useful for detection and identification of Brucella spp. A confirmed diagnosis requires isolation of Brucella from clinical specimens that is the most sensitive method in the acute and sub-acute phases of the diseases. On the other hand, molecular diagnostic techniques are more sensitive and more specific than serological techniques, especially in chronic localized cases because of antigenic cross-reactions or antibody titers lower than 160. Until now different Brucella specific sequences like BCSP 31, IS711 and 16SrRNA have been amplified for detection of Brucella spp. In this study, the sensitivity and specificity of The B4-B5 primers and IS711 designed primers were evaluated for detection of of Brucella Spp. in the clinical samples. Results : Amplification of extracted DNA from serum of 49 suspected patients were tested with two sets of specific primers. The BCSP31 amplicon was 223 bp and all the 49 (100%) serum specimens were positive by B4-B5 primers, including 4 cases with negative 2ME test result. The designed IS711 primers amplified the IS711 product with 448 bp length and 46 of 49 (93.87%) cases were positive. The sensitivity of the applied primers (B4-B5 and IS711) was evaluated by using the serial dilutions of extracted purified DNA molecules of B. melitensis and B. abortus . The B4-B5 primers can detect the least number of both B. melitensis and B. abortus , 0.1 CFU/reaction. However, the designed IS711 set is able to detect 10 CFU/reaction. The B4-B5 primer and IS711 designed primer recognized 100% (49/49) and 94% (46/49) of the cases, respectively. Conclusion: This study indicated that the sensitivity of B4-B5 primer is 100%, while the sensitivity of the designed primer of IS711 is 94%. The laboratory experiment revealed that designed IS711 set is 1×10 2 times more sensitive than sensitivity of the other experiments for detection of IS711 target sequence in the specimens.

scholarly journals Molecular diagnostics of infectious diseases

1997 ◽  
Vol 43 (11) ◽  
pp. 2021-2038 ◽  
Author(s):  
Yi-Wei Tang ◽  
Gary W Procop ◽  
David H Persing
Keyword(s):  
Nucleic Acid ◽  
Infectious Diseases ◽  
Direct Detection ◽  
Clinical Chemistry ◽  
Gene Mutations ◽  
Diagnostic Techniques ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
Molecular Diagnostic Techniques ◽  
Plasmid Profiling

Abstract Over the past several years, the development and application of molecular diagnostic techniques has initiated a revolution in the diagnosis and monitoring of infectious diseases. Microbial phenotypic characteristics, such as protein, bacteriophage, and chromatographic profiles, as well as biotyping and susceptibility testing, are used in most routine laboratories for identification and differentiation. Nucleic acid techniques, such as plasmid profiling, various methods for generating restriction fragment length polymorphisms, and the polymerase chain reaction (PCR), are making increasing inroads into clinical laboratories. PCR-based systems to detect the etiologic agents of disease directly from clinical samples, without the need for culture, have been useful in rapid detection of unculturable or fastidious microorganisms. Additionally, sequence analysis of amplified microbial DNA allows for identification and better characterization of the pathogen. Subspecies variation, identified by various techniques, has been shown to be important in the prognosis of certain diseases. Other important advances include the determination of viral load and the direct detection of genes or gene mutations responsible for drug resistance. Increased use of automation and user-friendly software makes these technologies more widely available. In all, the detection of infectious agents at the nucleic acid level represents a true synthesis of clinical chemistry and clinical microbiology techniques.

scholarly journals Are Nanobiosensors an Improved Solution for Diagnosis of Leishmania?

Pharmaceutics ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 491
Author(s):  
Sona Jain ◽  
Wanessa Santana ◽  
Silvio S. Dolabella ◽  
André L. S. Santos ◽  
Eliana B. Souto ◽  
...  
Keyword(s):  
Low Income ◽  
Neglected Tropical Diseases ◽  
Diagnostic Techniques ◽  
Cross Reactivity ◽  
Low Income Countries ◽  
Future Research ◽  
Immunological Tests ◽  
Diagnostic Approaches ◽  
Signal Improvement ◽  
Adequate Management

Leishmaniasis is one of the deadliest neglected tropical diseases affecting 12–15 million people worldwide, especially in middle- and low-income countries. Rapid and accurate diagnosis of the disease is important for its adequate management and treatment. Several techniques are available for the diagnosis of leishmaniasis. Among these, parasitological and immunological tests are most widely used. However, in most cases, the utilized diagnostic techniques are not good enough, showing cross-reactivity and reduced accuracy. In recent years, many new methods have been reported with potential for improved diagnosis. This review focuses on the diagnosis of Leishmania exploring the biosensors and nanotechnology-based options for their detection. New developments including the use of nanomaterials as fluorophores, fluorescence quenchers as reducing agents and as dendrimers for signal improvement and amplification, together with the use of aptamers to replace antibodies are described. Future research opportunities to overcome the current limitations on the available diagnostic approaches are also discussed.

scholarly journals Moraxella catarrhalis: from Emerging to Established Pathogen

2002 ◽  
Vol 15 (1) ◽  
pp. 125-144 ◽  
Author(s):  
Cees M. Verduin ◽  
Cees Hol ◽  
André Fleer ◽  
Hans van Dijk ◽  
Alex van Belkum
Keyword(s):  
Moraxella Catarrhalis ◽  
Current Knowledge ◽  
Bacterial Species ◽  
Bacterial Pathogen ◽  
Diagnostic Techniques ◽  
Human Infection ◽  
Molecular Diagnostic ◽  
Molecular Diagnostic Techniques ◽  
Pathogenic Potential ◽  
The Past

SUMMARY Moraxella catarrhalis (formerly known as Branhamella catarrhalis) has emerged as a significant bacterial pathogen of humans over the past two decades. During this period, microbiological and molecular diagnostic techniques have been developed and improved for M. catarrhalis, allowing the adequate determination and taxonomic positioning of this pathogen. Over the same period, studies have revealed its involvement in respiratory (e.g., sinusitis, otitis media, bronchitis, and pneumonia) and ocular infections in children and in laryngitis, bronchitis, and pneumonia in adults. The development of (molecular) epidemiological tools has enabled the national and international distribution of M. catarrhalis strains to be established, and has allowed the monitoring of nosocomial infections and the dynamics of carriage. Indeed, such monitoring has revealed an increasing number of Β-lactamase-positive M. catarrhalis isolates (now well above 90%), underscoring the pathogenic potential of this organism. Although a number of putative M. catarrhalis virulence factors have been identified and described in detail, their relationship to actual bacterial adhesion, invasion, complement resistance, etc. (and ultimately their role in infection and immunity), has been established in a only few cases. In the past 10 years, various animal models for the study of M. catarrhalis pathogenicity have been described, although not all of these models are equally suitable for the study of human infection. Techniques involving the molecular manipulation of M. catarrhalis genes and antigens are also advancing our knowledge of the host response to and pathogenesis of this bacterial species in humans, as well as providing insights into possible vaccine candidates. This review aims to outline our current knowledge of M. catarrhalis, an organism that has evolved from an emerging to a well-established human pathogen.

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