detection

Canine Distemper have become a major concern within the veterinary clinical work. Thus, the appearance of many cases of canine distemper in adult animals with their vaccination plan up to date has alarmed veterinarians. Many cases of canine distemper have become a major concern within the clinical veterinary task. The purpose of this work was to detect the gene of hemagglutinin of the virus canine distemper, by using the Polymerase Chain Reaction associated with Reverse transcription (RT-PCR), as confirmation of the clinical diagnosis of disease. For this, peripheral blood samples of animals were used clinically patients and were grouped according to their date of extraction and using vaccines commercial as control. The results allow to demonstrate a high sensitivity of the technique, besides allowing the use of samples up to seven days of storage at 4°C, despite the fragility of RNA viral. Detection of canine distemper virus haemagglutinin gene in field samples and its high sensitivity, suggests studying its use as a diagnostic tool complementary to the clinical diagnosis of canine distemper in our country.

Worldwide, Canine Distemper Virus (CDV) infection is a highly prevalent disease with high morbidity and mortality. CDV causes a multisystemic disease in a wide range of hosts including 9 families of mammals among them some primates, cetaceans and numerous carnivores. It presents a high tropism for lymphoid, neurological and epithelial tissue, leading to an infection of almost all systems, so the clinical signs observed are very varied. The diagnosis is made based on the clinical presentation of the disease, which considers a variety of signs and must be confirmed by a laboratory diagnostic method. The molecular technique called Reverse Transcription Polymerase Chain Reaction (RT-PCR) has been used to characterize viral strains based on the basis of genetic differences on the hemagglutinin (H) gene of CDV has allowed the identification of 14 circulating lineages in the world. Two lineages, namely the America-1 and the Europe-1/South America-1 have been described in Chile. The goal of this work was to implement a multiplex RT-PCR protocol, which was built on the in silico design of primers based on the H gene nucleotide sequences stored in the Genbank® database. This method was capable of detecting the previously described two circulating genetic lineages of CDV in a differential way providing a supporting diagnostic tool for epidemiological studies in the country. These results suggest that the primers described here are extremely selective for the above-mentioned lineages. In addition, our initial screening indicated that most analyzed clinical samples corresponded to the America-1 lineage, stressing the need for a continuous surveillance in order to properly address the prevalence of both lineages in Chile.

Oral cancer has a tendency to be detected at late stage which is detrimental to the patients because of its high mortality and morbidity rates (survival rate 15-50% [1]). The incidence of oral cancer worldwide is approximately 3% of all malignancies, thus creating a significant worldwide health problem [2].

The study was conducted in Ilu and Waliso districs of South West Shoa Zone to investigate the perception of dairy cattle producers on oestrus synchronization and mass artificial insemination services. A total of 122 respondents (38 from Ilu and 84 from Waliso districts) owning at least one lactating cow and participated in synchronization program were randomly selected and interviewed individually by using semi- structured questionnaire. Both primary and secondary data were used to generate reliable information on the intended topic. All collected data were analyzed using SPSS statistical package version 20 and the output was presented by descriptive statistics such as percentage and mean values in inform of graphs. Most of the respondents (67.15%) indicated that their satisfaction level towards synchronization and mass artificial insemination was low. Only few of them reported being satisfied as (medium-13.45%, good - 12.35% and very good - 7.05%). This might be because of shortage of feed, silent heat, poor performance of the inseminator and low awareness of farmers on the technology. From the study it was also noted that the overall percentage (26.22%) of calving rate to oestrus synchronization and mass artificial insemination was low. This might be due to heat detection problem (36%); A.I technician efficiency (29.25%), absence of A.I technician (23.9%) and distance of A.I center (10.25%). Therefore, the skill and knowledge based training should be given for both the farmers and implementers to enhance perception and adoption of the technology.

Botulism is the disease caused by botulinum neurotoxins. It is produced by an obligate anaerobic bacteria called Clostridium botulinum. There is no immuno-detection system available in the world for the detection of C. botulinum. Secretory proteins of cooked meat media grown C. botulinum type B were extracted by TCA precipitation method. Polyclonal antibodies were generated against secretory proteins. Cytokine profiling of secretory proteins were done. An immunodetection system was developed to detect the C. botulinum type B using Secretory proteins of C. botulinum type B.

Typhoid fever is a systemic infection caused by Salmonella enterica serotype typhi. It is of major concern in tropical regions of the world. Highest episodes of typhoid fever occur in Asia i.e.93%. Early diagnosis of the disease is mandatory to lower the mortality rate associated with it as well as to prevent the emergence of antimicrobial drug resistance by Salmonella typhi. Research work was conducted in Immunology Department of the Children’s Hospital, Lahore for the period of one year including a total of 60 patients suspected of having typhoid fever. Serum samples of these patients were tested for typhidot IgG and IgM antibodies as well as for the antibodies against TO and TH antigens using Widal test. Of the total 60 patients, 10 (16.7%) were positive for both typhidot IgG and IgM, 16 (26.7%) were positive for typhidot IgM, 3 (5%) were Positive for typhidot IgG and 31 (51.66%) were negative for both typhidot IgG and IgM. Reading the results of Widal test, 8 (13.33%) were positive for Widal TO and TH antigens, 3 (5%) were positive for Widal TO antigen, 19 (31.7%) were positive for Widal TH antigen and 30 (50%) were negative for Widal TO and TH antigens. IgM is positive at the early stage of acute typhoid fever, IgM along with IgG positive means the middle stage of acute illness. The detection of only IgG cannot discriminate between acute and convalescent phases as it can stay in the serum for at least 2 years or more. The typhidot test is much helpful for the rapid diagnosis of typhoid fever as compared to Widal test which is still being used in some set ups in poor countries, although has become mostly obsolete. By testing the rise of IgM and IgG antibodies against Salmonella typhi, we can detect the infection at early and late stages, respectively

Isocitrate dehydrogenase (IDH) mutations are a common event in secondary glioblastoma multiforme and lower-grade adult infiltrative astrocytomas and independently confer a better prognosis [1,2]. These are highly conserved mutations during glioma progression and thus also a useful diagnostic marker amenable to modern molecular sequencing methods. These mutations can even be detected in sites distant from the primary tumour. We use an illustrative case of a patient with radiologically suspected recurrent astrocytoma and negative histology, but positive IDH-mutated tumour DNA detected within CSF. Our results demonstrated the usefulness of liquid biopsy for recurrent glioma within the context of equivocal or negative histopathological results, whilst also showing the ability to detect a de-novo IDH-2 mutation not present in the previous resection. Building on this ‘proof-of-concept’ result, we also take the opportunity to briefly review the current literature describing the various liquid biopsy substrates available to diagnose infiltrative gliomas, namely the study of circulating tumour DNA, circulating tumour cells, and extracellular vesicles. We outline the current challenges and prospects of liquid biopsies in these tumours and suggest that more studies are required to overcome these challenges and harness the potential benefits of liquid biopsies in guiding our management of gliomas

Introduction: Chronic endometritis (CE) is a common cause of infertility in asymptomatic patients and its diagnosis and treatments improved assisted reproduction technique outcome in most of the specialized centers. Diagnosis of CE in endometrial biopsy by Hematoxylin and Eosin (H&E) stain is hard to identify chronic inflammatory cells from the stroma and the use of plasma cells-specific stains is helpful. Aim of the work: Evaluation of the use of CD138 in the identification of plasma cells in endometrial biopsy of patients with previous IVF trial failure. Material and methods: Hysteroscopic and curettage endometrial biopsies from fifty-five females with previous IVF trial failure were stained with H&E and CD138 immunostaining for detection of plasma cells. Results: Plasma cells were identified in 52.7% of cases by H&E and in 6/55 by CD138 immunostaining. CD138 is more sensitive in detecting plasma cells in endometrial biopsy than H&E stain. There was a significant statistical correlation between CE and abnormal uterine bleeding, abortion and primary infertility (p > 0.5). Conclusion: Diagnosis of CE is helpful in infertility patients with IVF trial failure to improve the outcome of the maneuver. CD138 is more sensitive for plasma cells specially in endometrial biopsies than H&E.

Objective: To observe the predominance of fetal anomalies in pregnant women in a multi-centric setting. Methods: This prospective observational study included 20225 pregnant women who came for antenatal care in University Hospital and fetal medicine units from 2016 to 2019. Fetal anatomical scanning was done for all participants. Results: One hundred eighty-three cases had fetal congenital anomalies, yielding a prevalence of around 0.9%. Third of cases had positive consanguinity, this increased in cases of skeletal and thoracic anomalies. The presence of past history of anomalies was evident in 8.2% mostly with skeletal and heart anomalies. History of drug intake was only verified in 1.6% of cases. Sixty-three women out of 183 (34.4%) were diagnosed to have anomalies in fetal nervous system. Conclusion: Prenatal diagnosis are recommended for early detection of congenital anomalies and counselling.

The industrial production and use of chromium have grown considerably during the past five decades. Abundances of the chromium isotopes in terrestrial samples are identical to 0.01%. Among the dominant species of chromium, the trivalent form widely occurs in nature in chromite ores which is extremely immobilized especially in water bodies. Samples were mixtures of separated chromium isotopes and the calibration was made with the same species as those used in the measurements. The method had simplified the conversion of the ores to chromyl fluoride since the element could be readily separated as lead chromate from the leaching of chromite-sodium peroxide fusions. Isotope assay of chromyl fluoride under certain conditions was measured and the measurements of chromium isotopic anomalies ratios and isotope abundance of the chromite ores have been assessed. These provided sufficient quantitative mass spectrometric data, which were analyzed to calculate the abundance and the mean atomic mass of the questioned isotopes. Based on the high mass spectroscopy stability and the correction factors, the results were of good precision (incl. negligible systematic errors normally associated to inter-laboratory discrepancies) and the Cr isotopes availability (52Cr > 53Cr > 50Cr > 54Cr) was in conjunction with other classical tools such as oxygen isotopes. This paper is important for paleoecological, environmental, archeological, forensic, and nuclear researchers.

Background: The use of brain magnetic resonance imaging (MRI) for evaluation of neurological disorders has increased in the past two decades. This has led to an increased detection of incidental findings on brain MRI. The most common of these asymptomatic abnormalities are white matter lesions that are interpreted as demyelinating based on radiological criteria. However, in the absence of associated clinical symptoms suggestive of multiple sclerosis (MS), a definite diagnosis of MS can’t be made in patients with these incidental white matter lesions. These patients are diagnosed as CIS (clinically isolated syndrome) and RIS (radiologically isolated syndrome).Using the revised McDonald criteria now allows some patients who would have been diagnosed with CIS to be diagnosed as having MS before a second episode. Method: Sixty one patients, 40 females and 21 males, age ranged between 15 years and 58 years, were included in our study. In addition to a detailed medical and neurological history and examination, CSF and blood analysis for oligoclonal bands and IgG index were performed for all patients. Result: 41 patients had positive oligoclonal bands and IgG index. After clinical, MRI results and laboratory results 44 (72.1%) were diagnosed CIS and 17 (27.9%) were RIS. Conclusion: Diagnosis of MS not depend only on MRI finding but need clinical and laboratory work up including CSF and blood analysis for oligoclonal bands and IgG index to confirm diagnosis.

Introduction: Hypertensive crisis (HC) is recognized consequence of inadequate blood pressure (BP) control. A hypertensive crisis is further divided into hypertensive emergency (HT-E) and hypertensive urgency (HT-U). Method: Using a cross-sectional hospital-based study design, patients who had been diagnosed as having HC between January and October 2017 were consecutively recruited in the study. The criteria proposed by the Seventh Joint National Committee were used for the definition of HC. Result: A total of 81 (.81%) patients newly diagnosed as having HC were enrolled in the study. Of these patients, 50 (61.7 %) patients met criteria for HT-E, while 31 (38.3%) patients had HT-U. Renal impairment (16%), stroke (30.8%), acute coronary syndrome (13.6%) and heart failure (22.2%) were predominant complications associated with HT-E. Out of 81 study subjects, 13 (16%) patients died. Although there was no significant difference in residence, history of smoking, Diabetes mellitus and history of alcohol consumption between groups, old age (P=.o22), male gender(.046), history of hypertension(.007), history of non-governmental employee(.003), poor compliance (p=.002) and high case fatality rate (p=.041) were significantly associated with hypertensive emergency (HT-E). Conclusion: This study showed that HT-E has high case fatality rate among patients admitted with hypertensive crisis at kassala teaching Hospital. Therefore early detection of hypertension and appropriate management are the main stay for reducing morbidity and mortality among patients with hypertensive crisis.

Background: Glaucoma is a multi-factorial optic neuropathy characterized by a loss of retinal ganglion cells with subsequent loss of the retinal nerve fibers ultimately resulting in visual impairment. The macula region has a high density of retinal ganglion cells thereby being a likely region to detect early cell loss .Since glaucoma affects mainly the inner layers of the retina, Ganglion Cell Complex (GCC) mapping can help to detect glaucomatous damage early as compared to the total retinal thickness. Purpose: To map GCC thickness and average Macular Retinal (MR) thickness with high-speed Fourier-Domain Optical Coherence Tomography (FD-OCT) and correlate it with the Retinal Nerve fiber layer (RNFL) thickness in preperimetric glaucoma. Design: Observational cross-sectional study. Methods: Forty four eyes diagnosed as preperimetric glaucoma were studied. GCC, MR thickness and RNFL thickness was mapped using the RTVue FD-OCT system. The GCC thickness map, the deviation map and the significance map were obtained in all cases. Average GCC thickness and MR thickness were correlated with the RNFL thickness. Results: Average GCC of patients was 85.99±6.9 µm. There was GCC loss in 35 (87.5%) eyes which correlated well with areas of RNFL loss (r=0.408, p<0.001). Nine (22.5%) eyes were seen to have decreased MR thickness. GCC loss correlated well with the loss of average RNFL thickness and MR thickness. Further GCC loss was also seen in 23 (74.19 %) eyes with a normal MR thickness. Conclusion: GCC analysis may prove to be a robust diagnostic parameter and is complementary to RNFL analysis in preperimetric glaucoma.

Obesity is a chronic and metabolic disease with a high increasing prevalence worldwide. It has multifactorial pathogenesis including genetic and behavioral factors [1-5]. Overweight and obesity have been defined and classified by the World Health Organization (WHO) and the National Institutes of Health (NIH) [2,3]. A person with a normal weight has Body Mass Index (BMI) of 18.5-24.9. A person with a BMI under 18.5 is called underweight. An adult having a BMI of 25-29.9 is overweight and pre-obese. Class 1 obesity is defined as a BMI between 30.00-34.99. Class 2 (Severe) Obesity is to have a BMI between 35.00-39.99. Morbid (Extreme, Class 3) obesity is to have a BMI over 40 [1-5]. Obesity is significantly associated with enhanced morbidity and mortality rates. It has also various economic, medical and psychological effects and causes health problems including many systemic diseases, economic costs and burdens, social and occupational stigmatization and discrimination and productivity loss [4-6]. Obesity carries the increased risk of development of many systemic and chronic diseases, including sleep apnea, depression, insulin resistance, Type 2 (adult-onset) diabetes, Gout and related arthritis, degenerative arthritis, hypertension, dyslipidemia, heart disease such as myocardial infarction, congestive heart failure, or coronary artery disease, polycystic ovary syndrome and reproductive disorders, Pickwickian syndrome (obesity, red face and hypoventilation), metabolic syndrome, non-alcoholic fatty liver disease, cholecystitis, cerebrovascular accident, colonic and renal cancer, rectal and prostatic cancer in males, and gallbladder, uterus and breast cancer in females [6-12]. In recent years, some publications reported that obesity has been strongly associated with some ocular diseases including age-related cataract and maculopathy, glaucoma, and diabetic retinopathy [13-16]. The recent reports demonstrated that the central corneal thickness and intraocular pressure were increased while as mean thickness of RNFL and retinal ganglion cell and choroidal thickness (CT) were decreased in the morbidly obese subjects [17-19]. However, another study has reported that CT increased in obese children [20]. On the other hand, a recent study reported that all values of the specific tests used to evaluate the ocular surface were within the normal range [21]. In some experimental studies, it has been demonstrated that obesity may cause retinal degeneration [22,23]. Additionally, in a past meeting presentation, it has been speculated that keratoconus is associated with severe obesity [24]. Teorically, idiopathic intracranial hypertension, and papilledema may also be associated with obesity [25]. Obesity may be also a cause of mechanical eyelid abnormalities such as entropion [26]. However, further investigations are needed to detect the significant relationship between these diseases and obesity. On the other hand, the ocular surgeries of obese patients are difficult compared to normal weight-subjects. The posterior capsule rupture and vitreous loss may easily develop during cataract surgery of these patients because obese patients have an elevated vitreous pressure and operating table cannot often be lowered or surgeon’s chair cannot be elevated sufficiently to provide the clear viewing of the operating area and tissues. So, some different surgical manipulations such as standing phacoemulsification technique and reverse Trendelenburg position have been developed. Additionally, the standing vitrectomy technique has been used for vitreoretinal interventions in morbidly obese patients [27,28]. In conclusion, all obese subjects should be subjected to a completed ophthalmological examination and to relevant clinics for the detection of possible comorbidities and diseases

To determine the proportion of participants with elevated intraocular pressure and abnormal vertical cup disc ratio in a glaucoma screening event in a rural community in Nigeria. Methods: 242 eyes of 121 participants at a one-day screening programme were examined. Visual acuity was accessed using Snellen literate and illiterate charts, followed by a pen torch examination of the anterior segment. The posterior segment was accessed using a direct ophthalmoscope (Welch Allyn, USA). All participants had intraocular accessed using Keeler non contact tonometer and those with significant media opacity preventing view of the posterior pole had cycloplegic examination after dilatation with 1%Tropicamide drops. Data generated was expressed as percentages and means. Results: 121 participants (242 eyes) were examined. There were 42 males and 79 females giving a M:F ratio of 1:1.86. Age range of participants was 0-90years with a peak in the 5th decade. Mean age was 32 years. 79.3% of participants had normal visual acuity of 6/4 -6/18, while 9.5% had visual acuity worse than 6/60. 83% of eyes had intraocular pressure within normal level, <21mmHg, 12.4% within 21-30mmHg and 4.6% >30mmHg. 73.1% of eyes had Vertical Cup disc ratio (VCDR) <0.5, 10.7% had 0.5-0.7 and 6.6%>0.7. 9.6% of eyes could not be accessed due to media opacities. Conclusion: 6.6% of participants had suspicious discs and 4.6% participants had intraocular pressures above 30mmHg. Opportunistic screening for glaucoma remains one of the important modes of case detection for glaucoma in the developing climes and should be encouraged as a way to address the scourge of this blinding disease.

Microparticles (MPs) are considered important diagnostic biological markers in many diseases with promising predictive value. There are several methods that currently used for the detection of number and characterization of structure and features of MPs. Therefore, the MP detection methods have been remained pretty costly and time consuming. The review is depicted the perspectives to use coupling methods for MP measurement and structure assay. Indeed, there is large body evidence regarding that the combination of atomic force microscopy or coupling nanoparticle tracking analysis (NTA) with microbeads, plasmon resonance method and fluorescence quantum dots could exhibit much more accurate ability to detect both number and structure of MPs when compared with traditional flow cytometry and fluorescent microscopy. Whether several combined methods would be useful for advanced MP detection is not fully clear, while it is extremely promising.

Post-translational modification (PTM) refers to the covalent and enzymatic modification of proteins during or after protein biosynthesis. In the protein biosynthesis process, the ribosomal mRNA is translated into polypeptide chains, which may further undergo PTM to form the product of mature protein [1]. PTM is a common biological mechanism of both eukaryotic and prokaryotic organisms, which regulates the protein functions, the proteolytic cleavage of regulatory subunits or the degradation of entire proteins and affects all aspects of cellular life. The PTM of a protein can also determine the cell signaling state, turnover, localization, and interactions with other proteins [2]. Therefore, the analysis of proteins and their PTMs are particularly important for the study of heart disease, cancer, neurodegenerative diseases and diabetes [3,4]. Although the characterization of PTMs gets invaluable insight into the cellular functions in etiological processes, there are still challenges. Technically, the major challenges in studying PTMs are the development of specific detection and purification methods.

Background: The proposal that MALDI-ToF mass spectrometry could be used as a direct, rapid and affordable diagnostic tool in clinical laboratory medicine has moved from a theoretical possibility to a reality for Microbiology. Several studies have proposed the application of this technology in obstetric and gynaecological evaluation of patients. In particular, we have proposed that the adoption of MALDI-ToF mass spectrometry in examination of maternal pregnancy urine samples for the detection of Downs syndrome. Methods: A retrospective collection of 20 Down Syndrome and 100 non-aneuploid pregnancy urines at 12 to 14 weeks gestation, collected in 2007-2008 from high risk pregnancy cohorts, were examined by MALDI-ToF mass spectrometry in the mass/charge range between 1000 and 100000 m/z. Normalisation of spectral data was defined using mass bins of 100 m/z expressed as a percentage of the total ion count of the mass spectra from 2000 to 11000 m/z. Of the ninety 100 m/z bins, forty-six were identified as m/z bins at which statistically significant differences in spectra occurred between Downs and control/non-aneuploid samples. Based on the differences and variance, for values at these bins, weighted scores of the probability of being Downs were assigned. Comparative algorithms consisting of various mass bins were tested for ability to distinguish Down syndrome from non-aneuploid pregnancy. Results: Although various algorithms could distinguish Downs from non-aneuploid controls, it was found that gestational age was a confounding factor and that if separated into gestational age matched cohorts the ability to distinguish the groups improved dramatically e.g. whilst a 19 bins algorithm separated 100% of Downs from non-aneuploid pregnancies for a 9% false positive rate in the mixed gestational ages group; a two bin algorithm distinguished 100% of Downs for a 6% false positive rate for the 12 weeks gestational age pregnancies. Conclusion: Normalised MALDI-ToF mass spectra, at 2000 to 11000 m/z, of maternal urine gives rise to gestational age specific screening tests algorithms for Downs’s syndrome.

The sequence-independent, single-primer amplification (SISPA) enables the random amplification of nucleic acids, allowing the detection and genome sequencing of different viral agents. This feature of SISPA method provides evidence for application of it in monitoring the presence of adventitious RNA viruses in cell cultures. We evaluated SISPA method for the detection of a challenge RNA virus representing adventitious agent in cell cultures. Besides, by optimizing the SISPA method in our laboratory, we found false-positive results on negative control lanes in electrophoresis gels. To investigate the sources of contamination, false-positive results of SISPA were cloned into Escherichia coli cells, sequenced, and phylogenetically analyzed. This data revealed that the SISPA method can be used as an adjunct method to confirm the absence of unexpected adventitious RNA viruses in cell cultures. The phylogenetic analysis of SISPA contaminant sequences showed that the false-positive results were caused by nucleic acid amplification of commercial cDNA synthesis kit reagents, probably tracing back to expression plasmids and host ribosomal sequences, used for the production of enzymes. Therefore, laboratories using random amplification methods must be constantly aware of the potentials of such contaminations, yielding false-positive results and background noise in the final NGS reads.

Venom has a very complex and exclusive nature which has been introduced by recent advances in omics technologists. These methods have revealed a new insight into venom studies as venomics. Envenoming by venomous animals is a global concern due to the distribution of important medical species around the world. Treatment of envenomed victims is dependent on accurate and fast identification of animal species with different detection methods. In recent years, new methods have been introduced based on molecular and immunological techniques. Precise diagnosis of species of venomous animals is an essential factor for treatment with specific antivenoms. Venomics and antivenomics data sets help in the selection of specific antivenoms or production of novel antivenoms with greater efficacies.