scholarly journals The Performance of Seven Molecular Methods for the Detection of PRRSV

2020 ◽  
Vol 70 (1) ◽  
pp. 51-57
Author(s):  
Vesna Milićević ◽  
Branislav Kureljušić ◽  
Ljubiša Veljović ◽  
Miroslav Valčić ◽  
Nataša Stević ◽  
...  
Keyword(s):  
Viral Disease ◽  
Diagnostic Techniques ◽  
Reproductive Failure ◽  
Accurate Diagnosis ◽  
Molecular Methods ◽  
Respiratory Disorders ◽  
Routine Diagnosis ◽  
Boar Semen ◽  
Indirect Losses ◽  
Important Disease

AbstractPorcine Reproductive and Respiratory Syndrome is a viral disease of swine characterized by reproductive failure of breeding animals and respiratory disorders in all categories. The first PRRS case in Serbia was recorded in 2001 after illegal import of boar semen. PRRS is economically the most important disease due to significant direct and indirect losses. Today, for routine diagnosis of PRRS in infected herds serological methods (ELISA) and molecular methods are used. Although modern diagnostic techniques are very robust, exceptional diversity of the viral strains is often the obstacle for an accurate diagnosis. To estimate the performance of seven different methods for PRRSV genome detection, twenty samples were used. However, none of the methods was able to detect all PRRSV strains. The best sensitivity was obtained by combining two methods. Until today, there is no absolutely accurate test which enables the detection of all circulating strains.

scholarly journals Isolation of Mucorales from biological environment and identification of Rhizopus among the isolates using PCRRFLP

2019 ◽  
Vol 21 (2) ◽  
pp. 98-103
Author(s):  
Mahboobeh Madani ◽  
Mohammadali Zia
Keyword(s):  
Reliable Method ◽  
Enzymatic Digestion ◽  
Accurate Diagnosis ◽  
Molecular Methods ◽  
Clinical Samples ◽  
Molecular Approaches ◽  
Pcr Products ◽  
Microscopic Studies ◽  
Biological Environment

Background and aims: Mucorales are fungi belonging to the category of Zygomycetes, found much in nature. Culture-based methods for clinical samples are often negative, difficult and time-consuming and mainly identify isolates to the genus level, and sometimes only as Mucorales. Therefore, applying fast and accurate diagnosis methods such as molecular approaches seems necessary. This study aims at isolating Mucorales for determination of Rhizopus genus between the isolates using molecular methods. Methods: In this descriptive observational study, a total of 500 samples were collected from air and different surfaces and inoculated on Sabouraud Dextrose Agar supplemented with chloramphenicol. Then, the fungi belonging to Mucorales were identified and their pure culture was provided. DNA extraction was done using extraction kit and the chloroform method. After amplification, the samples belonging to Mucorales were identified by observing 830 bp bands. For enzymatic digestion, enzyme BmgB1 was applied for identification of Rhizopus species by formation of 593 and 235 bp segments. Results: One hundred pure colonies belonging to Mucorales were identified using molecular methods and after enzymatic digestion, 21 isolates were determined as Rhizopus species. The sequencing of PCR products and macroscopic and microscopic studies confirmed the existence of R. stolonifera, R. oryzae and R. caespitosus in the samples. Conclusion: Generally, developing a reliable method for determining Zygomycete species can be a useful tool for better understanding of the epidemiology of mucoromycosis.

scholarly journals Dengue disease diagnosis: A puzzle to be solved

2015 ◽  
Vol 62 (4) ◽  
pp. 617-629 ◽  
Author(s):  
Jaime Castellanos
Keyword(s):  
Public Health ◽  
Dengue Virus ◽  
Point Of Care ◽  
Viral Disease ◽  
Disease Diagnosis ◽  
Diagnostic Techniques ◽  
Viral Isolation ◽  
Dengue Disease ◽  
Health Authorities ◽  
The Impact

<p>Dengue is an infection caused by dengue virus and is the most<br />important arthropod transmitted viral disease in the world,<br />causing near 100 million cases and 50 000 fatalities each year.<br />Health authorities believe that these numbers will grow in<br />coming years. In Colombia, almost 600 municipalities are in<br />regions with Aedes aegypti circulation, and the presence of four<br />dengue serotypes has been demonstrated. Despite the increasing<br />knowledge about disease pathogenesis and the dengue virus,<br />some technical or scientific difficulties with diagnosing dengue<br />remain, negatively affecting both public health surveillance<br />and the appropriate attention to patients in health settings and<br />hospitals. This paper reviews the principles and developments<br />of the current diagnostic techniques for dengue, pointing out<br />the difficulties with making accurate dengue diagnoses and case<br />confirmations in public health and specialized laboratories. The<br />principles and limitations of MAC-ELISA, IgG serology, viral<br />NS1 detection and viral isolation by cell culture are presented.<br />In addition, the review of immunochromatography techniques<br />(rapid diagnostic tests) that have been put forward to help the<br />point-of-care diagnosis is proposed. This paper is intended to<br />bring forward some points of view about the issues related to<br />dengue diagnosis and contribute to improve the discussion<br />surrounding the strategies and techniques needed for reducing<br />the impact of the disease and favoring its control.</p>

Aktualitások a sérült immunitású betegek cytomegalovirusinfekcióinak ellátásában. I. Epidemiológia és klinikai szempontok

2019 ◽  
Vol 160 (3) ◽  
pp. 83-92
Author(s):  
János Sinkó
Keyword(s):  
Viral Disease ◽  
Organ Damage ◽  
Diagnostic Techniques ◽  
Viral Reactivation ◽  
Cell Mediated Immunity ◽  
Solid Organ ◽  
Cytomegalovirus Reactivation ◽  
Vital Functions ◽  
Organ Specific ◽  
Clinically Significant

Abstract: Human cytomegalovirus is a DNA virus with a global prevalence of 40–100%. In humans, primary infection is followed by a lifelong latent persistent phase. Even in individuals developing adequate specific immunity, interactions with the resident virus do probably occur. Clinically significant problems, however, appear primarily in immunocompromised hosts. As a result of an impaired T-cell mediated immunity, viral reactivation as well as a viral disease resulting in organ damage can develop. Most severely affected are HIV positive persons in AIDS stage and individuals undergoing solid organ or stem cell transplantation. As vital functions and survival may adversely be affected by cytomegalovirus reactivation and disease, it is of paramount significance to evaluate evident risk factors. Viral reactivation and organ specific disease can be detected by several methods based on conventional and molecular biological and histological diagnostic techniques. An up-to-date management of affected patient groups requires a meticulous assessment of the possible risk for cytomegalovirus infection and development of an adequate antiviral strategy. Orv Hetil. 2019; 160(3): 83–92.

scholarly journals An Overview of the Molecular Methods in the Diagnosis of Gastrointestinal Infectious Diseases

2020 ◽  
Vol 2020 ◽  
pp. 1-13
Author(s):  
Muhammad Amjad
Keyword(s):  
Infectious Diseases ◽  
Diagnostic Techniques ◽  
Antigen Detection ◽  
Molecular Methods ◽  
Advantages And Disadvantages ◽  
Laboratory Setup ◽  
Detection And Identification ◽  
Intestinal Infections ◽  
Wide Range ◽  
Trained Staff

Gastrointestinal infectious diseases are very common worldwide and an important cause of morbidity and mortality, particularly in infants in developing countries. Diarrhea and other intestinal infections are caused by a wide range of bacteria, viruses, protozoa, and parasites. Conventional diagnosis of these infections is performed by culture, microscopy, and antigen detection immunoassays. The traditional culture and microscopy procedures are time-consuming, lack sensitivity, and require special laboratory setup and well-trained staff. However, based on the advancement in the molecular diagnostics and with the introduction of commercially available tests, traditional diagnostic techniques have been continuously replaced by these newer rapid antigen detection and molecular-based methods. This review summarizes and discusses the availability, advantages, and disadvantages of molecular methods in the detection and identification of human gastrointestinal pathogens.

scholarly journals Evaluation of a direct immunofluorescent antibody (difma) test using Leishmania genus - specific monoclonal antibody in the routine diagnosis of cutaneous leishmaniasis

1995 ◽  
Vol 28 (2) ◽  
pp. 99-103 ◽  
Author(s):  
Martha E. Chico ◽  
Ronald H. Guderian ◽  
Philip J. Cooper ◽  
Rodrigo Armijos ◽  
Max Grogl
Keyword(s):  
Monoclonal Antibody ◽  
Cutaneous Leishmaniasis ◽  
Diagnostic Techniques ◽  
Diagnostic Methods ◽  
Specific Monoclonal Antibody ◽  
Immunofluorescent Antibody ◽  
Routine Diagnosis ◽  
Highly Sensitive ◽  
Diagnostic Samples

A direct immunofluorescent antibody (DIFMA) test using a Leishmania genus- specific monoclonal antibody was evaluated in the routine diagnosis of cutaneous leishmaniasis (CL) in Ecuador. This test was compared with the standard diagnostic techniques of scrapings, culture and histology. Diagnostic samples were taken from a total of 90 active dermal ulcers from patients from areas of Ecuador known to be endemic for cutaneous leishmaniasis. DIFMA was positive in all lesions. It was shown to be significantly superior to standard diagnostic methods either alone or in combination. The sensitivity of DIFMA did not diminish with chronicity of lesions. This test proved to be extremely useful in the routine diagnosis of CL because it is highly sensitive, is easy to use and produces rapid results.

Clostridioides difficile laboratory diagnostic techniques: a comparative approach of rapid and molecular methods

2021 ◽  
Author(s):  
Thais Simões Camargo ◽  
Moacyr Silva Junior ◽  
Luis Fernando Aranha Camargo ◽  
Veronica Pivetta Biotto ◽  
André Mario Doi ◽  
...  
Keyword(s):  
Diagnostic Techniques ◽  
Molecular Methods ◽  
Comparative Approach ◽  
Clostridioides Difficile

scholarly journals Next-Generation Molecular Diagnostics Development by CRISPR/Cas Tool: Rapid Detection and Surveillance of Viral Disease Outbreaks

2020 ◽  
Vol 7 ◽  
Author(s):  
Sonal Srivastava ◽  
Dilip J. Upadhyay ◽  
Ashish Srivastava
Keyword(s):  
Influenza Virus ◽  
Virus Disease ◽  
Point Of Care ◽  
Disease Outbreaks ◽  
Viral Disease ◽  
H1n1 Influenza ◽  
Diagnostic Techniques ◽  
Molecular Diagnostic ◽  
Molecular Diagnostic Techniques ◽  
Next Generation

Virus disease spreads effortlessly mechanically or through minute insect vectors that are extremely challenging to avoid. Emergence and reemergence of new viruses such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), H1N1 influenza virus, avian influenza virus, dengue virus, Citrus tristeza virus, and Tomato yellow leaf curl virus have paralyzed the economy of many countries. The cure for major viral diseases is not feasible; however, early detection and surveillance of the disease can obstruct their spread. Therefore, advances in the field of virus diagnosis and the development of new point-of-care testing kits become necessary globally. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) is an emerging technology for gene editing and diagnostics development. Several rapid nucleic acid diagnostic kits have been developed and validated using Cas9, Cas12, and Cas13 proteins. This review summarizes the CRISPR/Cas-based next-generation molecular diagnostic techniques and portability of devices for field-based utilization.

Prospects and Application of Nanotechnology for Diagnosis of Tuberculosis in Livestock: A Review

2021 ◽  
Author(s):  
Archana Bharti ◽  
Yamini Verma ◽  
Amita Dubey ◽  
Madhu Swamy ◽  
Ajit Pratap Singh
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Human Population ◽  
Cost Effective ◽  
Diagnostic Techniques ◽  
Accurate Diagnosis ◽  
Biomolecular Interaction ◽  
Innovative Strategy ◽  
Diagnostic Use ◽  
Increased Sensitivity ◽  
Short Time

Tuberculosis (TB) is an infectious and contagious disease caused by various strains of Mycobacterium and the foremost leading cause of death of animal and human population worldwide. Currently available conventional and modern diagnostic techniques are useful but they are neither rapid nor cost effective for early and accurate diagnosis, represents a cornerstone to eradicate TB worldwide by 2030. To overcome this awful situation an innovative strategy i.e. nanotechnology is helpful for early diagnosis of tuberculosis in short time with increased sensitivity and specificity through nano sized biomolecular interaction viz. enhanced visualization of fluorescent signals, visual colorimetric signals of amplified DNA product, nanofluidic technology, nuclear magnetic resonance technology (NMR) and prototype miniaturized device etc. Nanotechnology has introduced new paradigms for molecular diagnostics. However, some constraints of nanodiagnostic tool still there that make obstacle in its worldwide diagnostic use for TB.

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