Real-time Transcriptase-polymerase Chain Reaction Research Articles

Study of MicroRNA-124 in Patients with Lupus Nephritis.

Lupus nephritis is associated with a six-fold increase in mortality compared with the general population. MicroRNAs studies revealed that increased MicroRNA -21 and MicroRNA -155 levels represent risk factors for active LN patients. MicroRNAs can be used as biomarkers in the diagnosis of clinical stages of LN. The present study aimed to determine the level of miR-124 in patients with lupus nephritis by reverse transcriptase real-time polymerase chain reaction compared to healthy control and correlate its levels with biochemical findings in those patients. The study was a case-control study that included fifty patients with lupus nephritis in addition to fifty healthy controls. Blood samples from the participants were subjected to the determination of serological markers of SLE. Moreover, real-time PCR was used for the determination of miR-124. The comparison of Micro-RNA124 between patients and control subjects revealed a statistically significant decrease in Micro-RNA124 in patients (1.193 ± 0.56) compared to the control (3.36 ± 0.50, p <0.001); the comparison of the level of MicroRNA 124 in the patients with different clinical and serological findings of SLE revealed a significant decrease in the level of MicroRNA 124 in patients with muscular findings (1.02 ± 0.5) compared to the patients with negative manifestations (1.47 ± 0.5, p =0.005) Conclusion: In the present study, a comparison of MicroRNA-124 in LN patients with different stages compared to normal control showed a statistically significant decrease in Micro-RNA124 in patients with lupus nephritis p <0.001 with significant correlation to the patients' different clinical and serological findings of SLE. Therefore, it may be used as a new noninvasive therapeutic approach to monitor response to therapy, predict relapses, and identify the degree of the activity of the disease or the progression to the chronic stage.

Effects of Coronavirus Disease 2019 on Prevalence of Acute Respiratory Viruses: Changes during the Pandemic.

The coronavirus disease 2019 (COVID-19) pandemic may have influenced the prevalence and seasonality of acute respiratory viral infections. The aim of the study was to investigate the prevalence of all viruses causing acute viral respiratory infections before and after social distancing measures were lifted. Cross-sectional study where outpatients and inpatients at Kyunghee University Hospital were examined. From January 2021 to December 2022, respiratory samples were analyzed using multiplex reverse transcriptase real-time polymerase chain reaction. Of 3953 samples obtained, 412 (10.42%) were positive for acute respiratory viral infection, and 502 viruses were detected. The number of viral infections increased from 184 in 2021 to 318 in 2022. Human metapneumovirus was detected from August to November 2022. Human bocavirus (HBoV) was frequently detected from April to June 2021; however, in 2022, HBoV was frequently detected from July to October. Human parainfluenza virus 3 was rarely detected after its initial frequent detection from October to December 2021 but was continuously observed after frequent detection in September 2022. Co-infection occurred in 78 (18.9%) cases. The most common combination of simultaneous infections was human rhinovirus-HBoV (n = 30, 38.5%). During the COVID-19 pandemic, the incidence of acute respiratory viral infection decreased significantly but increased in 2022 when measures were lifted. The prevalence and seasonality of respiratory viral infections have changed since the pandemic. Our findings contribute to the prediction of an effective response to changes in the prevalence of respiratory viruses.

Mosquito Community in Primate Captivity (Tarsius sp.) and its Potential as Transmitters of Zoonotic Mosquito-Borne Diseases.

By means of conservation, ectoparasites monitoring and surveillance especially mosquitoes in primate captivity become important. Mosquito is one of the ectoparasites which acts as a vector of various types of zoonotic diseases such as Dengue, Zika, Japanese encephalitis, Rift Valley fever, filariasis, and others. This study was aimed to determine the diversity of mosquito species, their fluctuations as well as the potential of mosquitoes as a Dengue virus (DENV) vector around the tarsier captivity in the animal conservation laboratory IPB Primate Research Center (IPB PRC). Mosquitoes were collected from February to April 2020 using light traps and sweep net every two hours from 06.00 pm to 06.00 am. Detection of the presence of Dengue virus (DENV) is carried out using Reverse Transcriptase Real-time Polymerase Chain Reaction (RT-qPCR) against Aedes albopictus and Armigeres subalbatus. The results showed that there were 4 species of mosquitoes caught around the tarsier captivity namely Aedes albopictus, Culex quinquefasciatus, Armigeres subalbatus, and Armigeres foliatus. The dominant mosquito species were Armigeres subalbatus (62.11%) and Aedes albopictus (41.61%). Detection of Dengue virus (DENV) serotypes 1, 2, 3, 4 in Aedes albopictus and Armigeres subalbatus gave negative results. The presence of mosquitoes that have the potential to carry zoonotic disease around the tarsier captivity in IPB PRC shows the potential for Mosquito-Borne Diseases to both tarsiers and human.

Subgingival Plaque as a Potential Reservoir for SARS-CoV-2 Virus in Patients Recovered from COVID-19 Infection: A Cross-sectional Study

Background Subgingival plaque biofilm is a potential harboring site for bacteria and viruses, serving as a source of entry into the bloodstream. The presence of Severe Acute Respiratory Syndrome Coronavirus 2 RNA (SARS-CoV-2 RNA) in subgingival plaque could be a possible source for cross infection due to aerosol contamination during dental procedures. Objectives The objective of this study was to investigate the presence of SARS-CoV-2 RNA in the subgingival plaque biofilm of patients who recovered from COVID-19 infection. Materials and Methods Ten subjects visiting the Outpatient clinic with a previous history of COVID-19 infection were recruited for this cross-sectional study. Subgingival plaque sample was collected from all four quadrants using a sterile curette. Nasopharyngeal and oropharyngeal swabs were taken from all the subjects. Reverse transcriptase real-time qualitative polymerase chain reaction (RT-PCR) was used to determine the presence of SARS-CoV-2 RNA using BAG SARS-CoV-2 RT-PCR kit targeting E gene, RdRp gene, and internal control. Results Ten participants (seven females and three males) with a previous history of COVID-19 infection were included in the study. Six participants previously presented with mild COVID-19 infection (uncomplicated upper respiratory tract infection, mild fever, cough, sore throat, nasal congestion, malaise, headache, without evidence of breathlessness or hypoxia), and four participants previously presented with moderate COVID-19 infection (dyspnea and or hypoxia, fever, cough, SpO2 90%–94% on room air, respiratory rate more or equal to 24 per minute) according to clinical management protocol for COVID-19, Government of India. The nasopharyngeal, oropharyngeal, and subgingival plaque samples tested negative for SARS-CoV-2 RNA. Conclusion The absence of the SARS-CoV-2 virus in the subgingival plaque sample of patients recovered from COVID-19 infection suggests that dental plaque could not be a potential reservoir of the virus. However, clinicians should take at most precautions while treating the patients recovered from COVID-19 infection and should anticipate a change in subgingival environment leading to alteration in periodontal disease progression.

Serum level of MicroRNA 15a and interleukin 6 as biomarkers for sepsis in critically Ill Patients

This study intended to evaluate the diagnostic efficacy of serum interleukin-6 (IL-6) and serum expression level of microRNA 15a (miRNA 15a) in sepsis patients admitted to the medical intensive care unit (ICU), and to correlate the results with the outcome of patients. This observational case-control study, was done from January 2022 to June 2022, included 75 adult ICU patients ≥18 years old, divided into 2 groups. Group 1: included 38 adults, critically ill sepsis patients and Group 2 (control group), included 37 adults, non-sepsis patients admitted to the ICU. Venous blood was withdrawn from cases and controls under aseptic conditions for assessment of serum IL-6 level by ELISA and serum expression level of miRNA 15a by quantitative reverse transcriptase real-time polymerase chain reaction (qRT-PCR). Patient demographic data in addition to their characteristics including site of infection, blood culture results, length of stay and outcome of patients were included in the study. miRNA 15a was significantly higher in the sepsis group with mean of 3.99±1.61 compared to the controls (1.03±0.03, p&lt; 0.001) while serum IL-6 levels were higher in the sepsis patients compared to the controls, however the difference did not reach statistical significance (p= 0.92). No difference was found in IL-6 and miRNA 15a levels between surviving and non-surviving sepsis patients, shocked and non-shocked sepsis patients. In conclusion, both IL-6 and miRNA 15a were up-regulated in ICU patients with sepsis. Both biomarkers did not show a significant difference regarding the outcome of patients. miRNA 15a could be considered a highly specific and sensitive marker in differentiating sepsis cases from controls, so it could be used as a diagnostic rather than a prognostic biomarker for sepsis.

Ellagic acid inhibits the formation of hypertrophic scars by suppressing TGF-β/Smad signaling pathway activity.

Hypertrophic scar (HS) is a benign fibroproliferative skin disease, which lacks the ideal treatment and drugs. Ellagic acid (EA) is a natural polyphenol that prevents fibroblasts from proliferating and migrating. This study aimed to determine the role of EA in HS formation and its possible mechanism by in vitro experiments. HS fibroblasts (HSFs) and normal fibroblasts (NFs) were separated from HS tissue and normal skin tissue, respectively. HSFs were treated with 10 and 50 μM EA to assess their effect on HS formation. In particular, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and scratch assay were used to detect the viability and migration ability of HSFs. Quantitative reverse transcriptase real-time polymerase chain reaction was used to measure the mRNA expression level of basic fibroblast growth factor (bFGF), extracellular matrix (ECM)-related gene collagen-I (COL-I), and fibronectin 1 (FN1) in HSFs. Finally, Western blot was utilized to measure the expression level of TGF-β/Smad signaling pathway-related proteins in HSFs. The viability of HSFs was significantly increased compared with NFs. 10 and 50 μM EA treatment markedly inhibition the cell viability and migration of HSFs. EA treatment upregulated the bFGF expression level and downregulated the COL-I and FN1 expression level in HSFs. In addition, p-Smad2, p-Smad3, and transforming growth factor (TGF)-β1 expression levels as well as p-Smad2/Smad2 and p-Smad3/Smad3 ratios remarkably decreased in HSFs after EA treatment. EA inhibited the formation of HSs by suppressing the viability and migration of HSFs and ECM deposition as well as by preventing the activation of TGF-β/Smad signaling.

Viral etiology of lower respiratory tract infections in adults in the pre-COVID-19 pandemic era: A cross-sectional study in a single center experience from Cameroon.

Respiratory viruses are responsible for the majority of lower respiratory tract infections (LRTIs) worldwide. However, there is a gap on the epidemiology of viral LRTIs in adults in sub-Saharan African countries. In Cameroon, like in other countries, the role of viral respiratory pathogens in the etiology of LRTIs in adults is helpful for clinical management. This study aimed to determine the viral aetiologies of LRTIs among hospitalized adults in a reference center for respiratory diseases in the town of Yaounde in Cameroon and its surroundings. A cross-sectional study was conducted from January 2017 to January 2018 at Jamot Hospital in Yaounde (Cameroon). Clinical and demographic information; BAL and sputa were collected from hospitalized patients meeting LRTI case definitions. The clinical samples were investigated for respiratory pathogens with a commercial Reverse Transcriptase Real-Time Polymerase Chain Reaction (RT-PCR) targeting 21 viruses, cultures for bacterial and fungal infections. The 77 included adult patients with LRTIs had an appropriate clinical sample for microbial investigations. A viral agent was detected in 22.1% (17/77) samples. The main viruses detected included rhinovirus (10/77), coronavirus (hCoV-OC43 and hCoV-229E), and influenza A virus (3/77 each). A concomitant viral and bacterial co-infection occurred in 7.8% of patients (6/77) while viral co-infection occurred in one patient (1.3%). No Severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) was detected in clinical samples. Most patients were under antimicrobials before getting diagnosed. Respiratory viruses account for 22.1% of LRTIs in hospitalized patients in this study. Despite prior antimicrobial therapy and delay, rhinovirus, coronavirus and influenza A virus were the most detected in patients in the pre-COVID-19 pandemic era in a single center experience from Cameroon.

Evaluation of the Abbott Panbio™ COVID-19 Ag Rapid Antigen Test for Asymptomatic Patients during the Omicron Wave

Rapid antigen testing (RAT) is a cost-effective and time-efficient method of identifying severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and therefore a critical part of infection control strategies. There is no published evidence evaluating the use of RAT during the Omicron wave of the COVID-19 pandemic for asymptomatic patients or its performance between waves. All patients presenting to an Emergency Department over a two-week period without COVID-19 symptoms were screened for SARS-CoV-2 using both the Abbott Panbio RAT as well as the gold standard reverse transcriptase real-time polymerase chain reaction (PCR). The Abbott Panbio RAT sensitivity was 13% (95% CI 0.028, 0.336) for asymptomatic patients. The use of this test in asymptomatic patients during the Omicron wave had a statistically significant reduction in sensitivity compared with two reports of the same test in previous waves (13% vs. 86%, p &lt; 0.0001; 13% vs. 83%, p &lt; 0.0001). As SARS-CoV-2 continues to mutate, the sensitivity of RATs are altered and needs to be continually re-evaluated for each variant of concern if they are to be used as part of an infection control strategy.

Congruence of radiological scoring systems used in COVID-19 pneumonia and effect of comorbid diseases on radiological features

Background/Aim: Several scoring systems have been developed to standardize radiological findings in patients with COVID-19 pneumonia. The most commonly used scoring systems in the radiological examination of COVID-19 are those of the North American Radiology Association (RSNA), British Thoracic Society (BTS), and COVID-19 Reporting and Data System (CO-RADS). However, the compatibility between these radiological scoring systems has not been evaluated before. Therefore, this study evaluated the radiological features of COVID-19 pneumonia and congruence between radiological scoring systems and determined the effect of comorbidities and demographic characteristics on radiological features and thoracic computed tomography (TCT) findings in the context of COVID-19. Methods: A retrospective cohort study was performed on patients attending our unit with a suspicion of COVID-19 who also had a positive real-time transcriptase-polymerase chain reaction (RT-PCR) test. All TCT images were subjected to the RSNA, BTS, and CO-RADS scoring systems. Demographic data such as age and gender, and comorbid conditions were recorded. Results: TCT showed peripheral, posterior, and bilateral involvement in 97.7%, 97.7%, and 87.6% of the patients, respectively. The most common TCT finding was ground glass appearance, which was found in 95.5% of the patients. The Charlson Comorbidity Index (CCI) score was found to have an impact on RSNA and BTS criteria (P=0.011 and P=0.014), while age, gender, and the presence of comorbidities such as cardiovascular disease (CVD), diabetes mellitus (DM), and chronic pulmonary disease (CPD) did not have such an effect (P&gt;0.05 for all). On the other hand, CCI scores and the presence of CPD had an association with CO-RADS, but there was no effect of age, gender, DM, and CVD (P=0.915 and P=0.730). Conclusion: TCT plays an important role in early management, isolation, and follow-up of patients with COVID-19 pneumonia. The radiological scoring systems were found to exhibit good compatibility, but comorbid conditions could have an impact on the assessment. Therefore, we conclude that these radiological assessment criteria are useful in the management and monitoring of such patients.

An Estimate of Time-Dependent Transmission of COVID-19 Pandemic in Katsina State from June to November 2021

This paper investigates the spread of COVID-19 pandemic in Katsina state from 23rd June to 27th November, 2021, using graphical tools to visually analyse infection over time and time-dependent reproduction number to quantify the transmissibility of the disease. Study data consist of diagnostic results of reverse transcriptase real-time quantitative polymerase chain reaction assays (RT-PCR) on nine hundred and eighty-nine nasal swab samples from suspected COVID-19 cases, collected from public and private health facilities across Local government areas of the State and analysed by the Molecular Laboratory of the Federal Medical Centre Katsina, recording a total of 137 positive cases. Our investigation revealed that, over the study period, COVID-19 transmission reaches its peak on 17th October 2021 (Epidemic Week 41), with a mean Reproduction number (Rt) of 3.22 and a standard deviation of 0.5976 (95% CI: 2.6193 - 3.8145), culminating in 19 new infections on Wednesday the 27th (Epidemic Week 43). The combined age groups 21-30 and 31-40 constituted the most affected cases with 29 (21.17%) and 19 (13.87%) positive cases, respectively. This was closely followed by age group 51-60 with 19 (13.87%) positive cases. On gender basis, 70 (51%) females tested positive compared to 67 (49%) males. We therefore conclude that for the third wave of COVID-19 pandemic in Katsina State, disease transmission reaches its peak in Epidemic Week 41 in October, then rapidly diminishes to Rt values of less than one, indicating that no new infections were expected by the end of November, with female gender and age groups in the range 21 to 40 years being most infected. We recommend that mitigation strategies that take into consideration features inherent in female gender and more pronounced in young (21 to 40 years) and middle-aged (51-60 years) adults should be adopted in case of future waves.

Telomerase Reverse Transcriptase and Telomerase RNA Component Gene Expression as a Novel Biomarkers for Alzheimer's disease.

Alzheimer's disease (AD) is a neurological, age-related condition that causes cognitive decline and memory loss; it induces dementia in the elderly. Telomerase is a reverse transcriptase ribonucleoprotein that adds nucleotides to the end of DNA. This study aimed to compare human telomerase reverse transcriptase (hTERT) and telomerase RNA component (TERC) expression in different phases of AD and healthy cohorts. Sixty participants were divided into 30 who had dementia and 30 who did not. After collecting blood samples, total RNAs were extracted from the plasma. Screening for hTERT and TERC gene expression was carried out by quantitative reverse transcriptase real-time polymerase chain reaction (RT-qPCR) using the relative quantification method to estimate the expression changes in hTERT and TERC. The RT-qPCR results show that hTERT and TERC gene expression was significantly down-regulated in Alzheimer's patients compared to the health subjects (P-value= <0.0001,0.005), respectively. The area under curve AUC was 0.773 for hTERT and 0.703 for TERC. The Mini-Mental State Examination scores revealed a significant difference between dementia and non-dementia subjects (P=<0.0001). We conclude down-regulations in both hTERT and TERC gene expression in AD patients, which supports our hypothesis that the telomerase expression gene in the blood of AD patients can serve as a non-invasive, early, and novel diagnostic marker of AD.

Diagnostic Value of Natural Killer Cells, CD56+ CD16+ Natural Killer Cells, NLRP3, and Lactate Dehydrogenase in Severe/Critical COVID-19: A Prospective Longitudinal Study According to the Severe/Critical COVID-19 Definitions.

Innate immunity, as the first line of defense of our immune system, plays a crucial role in defending against SARS-CoV-2 infection and also its immunopathogenesis. We aim to investigate the immune status of natural killer (NK) cells, natural killer T (NKT) cells, and NLRP3 gene expression in COVID-19 patient blood samples. The immunophenotype of NK cell subsets and NKT cells was detected by flow cytometry and the expression of NLRP3 gene assessed by reverse transcriptase real-time polymerase chain reaction in 44 COVID-19 patients and 20 healthy individuals. The percentage of most of NK cell subpopulation and NKT cells was significantly decreased in COVID-19 patients. The percentage of CD56dim CD16- NK cell subsets, and NLRP3 gene expression increased. The percentage of total NK cells, CD56+ CD16+ NK cells, and NLRP3 gene expression had acceptable sensitivity and specificity for assisting diagnosis of severe/critical COVID-19. O2 saturation% and lactate dehydrogenase levels showed valuable diagnostic value to identify critical cases. The declined NK and NKT cells in COVID-19 patients and enhanced NLRP3 gene expression were associated with disease severity. Total NK cells, CD56+ CD16+ NK cells, and NLRP3 gene expression might be used as meaningful indicators for assisting diagnosis of severe/critical COVID-19.

SARS-CoV-2 Viral Load Quantification, Clinical Findings and Outcomes in Children Seeking Emergency Department Care: Prospective Cohort Study.

We compared the perfomance of SARS-CoV-2 reverse transcriptase real-time polymerase chain reaction (RT-PCR) to droplet digital PCR (ddPCR). 95% and 40% of positive and negative RT-PCR specimens, respectively, were positive on ddPCR yielding sensitivities of 84% (95% CI: 74, 91) and 97% (95% CI: 89, 99), for RT-PCR and ddPCR, respectively. We found that SARS-CoV-2 RT-PCR testing in children has a concerning false-negative rate at lower nucleocapsid gene copy numbers.

Repeated transmission of SARS-CoV-2 in an overcrowded Irish emergency department elucidated by whole-genome sequencing

AimTo provide a detailed genomic-epidemiological description of a complex multi-ward SARS-CoV-2 outbreak, which originated in the crowded emergency department (ED) in our hospital during the third wave of the COVID-19 pandemic, and was elucidated promptly by local whole-genome sequencing (WGS).MethodsSARS-CoV-2 was detected by reverse transcriptase real-time polymerase chain reaction on viral RNA extracted from nasopharyngeal swabs. WGS was performed using an Oxford MinION Mk1C instrument following the ARTIC v3 sequencing protocol. High-quality consensus genomes were assembled with the artic-ncov2019 bioinformatics pipeline and viral phylogenetic trees were built, inferred by maximum-likelihood. Clusters were defined using a threshold of 0–1 single nucleotide polymorphisms (SNPs) between epidemiologically linked sequences.ResultsIn April 2021, outbreaks of COVID-19 were declared on two wards at University Hospital Limerick after 4 healthcare-associated SARS-CoV-2 infections were detected by post-admission surveillance testing. Contact tracing identified 12 further connected cases; all with direct or indirect links to the ED ‘COVID Zone’. All sequences were assigned to the Pangolin B.1.1.7 lineage by WGS, and SNP-level analysis revealed two distinct but simultaneous clusters of infections. Repeated transmission in the ED was demonstrated, involving patients accommodated on trolleys in crowded areas, resulting in multiple generations of infections across three inpatient hospital wards and subsequently to the local community. These findings informed mitigation efforts to prevent cross-transmission in the ED.ConclusionCross-transmission of SARS-CoV-2 occurred repeatedly in an overcrowded emergency department. Viral WGS elucidated complex viral transmission networks in our hospital and informed infection, prevention and control practice.

Inconspicuous post-mortem findings in rabbits from Switzerland naturally -infected with Rabbit Haemorrhagic -Disease Virus 2.

Rabbit Haemorrhagic Disease Virus 2 (RHDV-2) emerged in France in 2010. In Switzerland, RHDV-2 was first identified in 2015 and apparently has almost replaced the classical Rabbit Haemorrhagic Disease Virus (RHDV) by now. Like RHDV, RHDV-2 causes a viral hepatitis with a peracute course and an increased mortality rate within the rabbitry. RHDV infection causes consistent gross pathological findings, especially in the liver and respiratory tract. Reports about gross pathology for animals naturally infected with RHDV-2 is scarce. The present study analysed the anamnesis and necropsy reports of 35 rabbits examined during routine diagnostics between March 2015 and May 2017. A reverse transcriptase real-time polymerase chain reaction (RT-qPCR) specific for RHDV-2 and RHDV proved a total of 25 animals to be positive for RHDV-2, while none was positive for RHDV. Additionally, histological examinations were performed on liver, lung, and kidney of 18 rabbits that had tested positive by RHDV-2 RT-qPCR. The anamnestic report more often stated an increased mortality rate in RHDV-2 positive (16/18, 89 %) compared to RHDV-2 negative rabbits (3/9, 33 %). Gross pathology did not reveal any pathognomonic changes in RHDV-2 positive animals. Histologically, the liver showed the most severe lesions followed by lung and kidney. Animals positive for RHDV-2 frequently showed signs of gastro-intestinal disease (n = 5) and/or septicaemia (n = 6) masking possible indicators of an RHDV-2 infection, such as the rather unspecific findings of an enlarged spleen or an enlarged, friable, tan-coloured liver. The authors want to raise awareness among clinicians and pathologists that in case of sudden death in commercial or pet rabbits, RHDV-2 needs to be considered as differential diagnosis and should be confirmed by laboratory diagnosis.

The effect of hypertension and antihypertensive therapies on the course of COVID-19 infection: Turkish national health system data

ABSTRACT Aim The effect of hypertension (HT) and antihypertensive therapies such as renin-angiotensin-aldosterone system (RAAS) blockers on the disease course in COVID-19 patients is controversial. The purpose of this study was to evaluate the effect of HT and antihypertensive therapies on the course of COVID-19 disease. Method The age, sex, comorbid diseases, and antihypertensive therapies of 132,790 patients with positive COVID-19 real-time transcriptase polymerase chain reaction (RT-PCR) tests in the Turkish Health Ministry National COVID-19 database between 11 March and 31 May 2020, were examined and analyzed. Results Forty-one percent of the 132,790 patients in this study (median age: 40, 47.3% female) were hospitalized for treatment, and 4.5% were followed-up in the intensive care unit (ICU). The most frequent comorbid disease, at 19.5%, was HT (n = 25,863). Mortality was determined in 4.9% of HT patients and 1.9% of non-HT patients (p < .001). HT, age, and male gender emerged as independent predictors of hospitalization and admission to the ICU, while HT was not a predictor of mortality. In addition, no adverse effect of any antihypertensive treatment, including RAAS inhibitors, on mortality was detected. Conclusion Based on Turkish national data, HT is common in COVID-19 patients, but does not appear to be an independent predictor of mortality, and no adverse effect of RAAS inhibitors on COVID-19-related mortality was observed.

A Pre- and Within-Pandemic Survey of SARS-CoV-2 RNA in Saliva Swabs from Stray Cats in Switzerland.

Cats have been shown to be highly susceptible to SARS-CoV-2, and transmission within the species has been demonstrated experimentally. In cats undergoing natural SARS-CoV-2 infections, human-to-animal transmission was mostly suspected. It can be postulated that, in stray cats with no or only minimal contact with humans, SARS-CoV-2 may pose a minor risk. The current study investigated the prevalence of active SARS-CoV-2 infections in Swiss stray cats using quantitative reverse transcriptase real-time polymerase chain reaction (RT-qPCR). Saliva swabs from 1405 stray cats were collected in 14 Swiss cantons. The animals were sampled between February 2019 and February 2020 (pre-COVID-19 cohort: 523 cats) and between February 2020 and August 2021 (COVID-19 cohort: 882 cats). All the samples were tested by RT-qPCR, amplifying the envelope (E) gene and, in case of positive or inconclusive results, the RNA-dependent RNA polymerase (RdRp) gene of SARS-CoV-2. No SARS-CoV-2 viral RNA could be detected in any of the tested saliva swab samples. Our findings support the assumption that SARS-CoV-2 infections in stray cats are not highly prevalent in Switzerland. Nevertheless, the monitoring of stray cats and other susceptible animal species is necessary, since the “One Health” approach has been recognized as being essential to successfully fight the COVID-19 pandemic.

A Molecular and Epidemiological Description of a Severe Porcine Reproductive and Respiratory Syndrome Outbreak in a Commercial Swine Production System in Russia.

Porcine reproductive and respiratory syndrome (PRRS) is an economically devastating disease of swine in many parts of the world. Porcine reproductive and respiratory syndrome virus (PRRSV) type 1 is endemic in Europe, and prevalence of the subtypes differ spatially. In this study, we investigated a severe PRRS outbreak reported in 30 farms located in eastern Russia that belong to a large swine production company in the region that was also experiencing a pseudorabies outbreak in the system. Data included 28 ORF5 sequences from samples across 18 of the 25 infected sites, reverse transcriptase real-time polymerase chain reaction (RT-qPCR) results from diagnostic testing, reports of clinical signs, and animal movement records. We observed that the outbreak was due to two distinct variants of wildtype PRRSV type 1 subtype 1 with an average genetic distance of 15%. Results suggest that the wildtype PRRSV variants were introduced into the region around 2019, before affecting this production system (i.e., sow farms, nurseries, and finisher farms). Clinical signs did not differ between the variants, but they did differ by stage of pig production. Biosecurity lapses, including movement of animals from infected farms contributed to disease spread.

A review on newer detection technologies for the diagnosis of COVID-19 infection

COVID-19 is a sickness brought about by coronavirus responsible for causing simple to extreme complications in people. COVID-19 first case was seen in Wuhan, Hubei Province, China, on December, 2019. The World Health Organization (WHO) to pronounce it as a worldwide pandemic on March 11, 2020, as the pandemic has spread quickly all through the world. Regardless of extensive endeavors made to contain the infection, the infection has proceeded with its pervasiveness in numerous nations with changing levels of clinical signs. Henceforth, in this report, we discuss the various strategies, for example, serological and nucleic acid-based procedures which are accessible for the determination and successful counteraction of coronavirus. With expanding the rate of coronavirus cases, the precise and early identification of the COVID-19 is the need of great importance for viable avoidance with treatment and just as to check its spread. Reverse transcriptase-real time polymerase chain reaction (PCR) measures are viewed as the highest quality level for the early identification of infection. This diagnostic technique is being utilized worldwide with recommendations from WHO and Center for Disease Control and Prevention. Reverse transcriptase real-time quantitative PCR (RT-qPCR) is being done compulsory before any medical procedures and major surgeries for early detection, prevention, and management in due time course. Rapid antigen test is also a screening test used widely in hospitals for screening of COVID-19 and before the admission in hospitals. Other nucleic acid amplification test widely done for the detection of COVID-19 are RT-qPCR, next-generation sequencing, clustered regularly interspaced short pallindromic repeats, reverse transcription -loop-mediated isothermal amplification, droplet digital PCR. Some immunological tests are lateral flow, ELISA.

Incidence of Non Amplification of N Gene of SARS-CoV-2 using Commercially Available Real Time Polymerase Chain Reaction Kit and its Comparison with Two Other Different Kits

Introduction: Novel Coronavirus Disease 2019 (COVID-19) caused by the Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) has resulted in an unprecedented global pandemic. Real Time Polymerase Chain Reaction (RT-PCR) tests are being used in the diagnosis of COVID-19 worldwide however mutations in the SARS-CoV-2 genome have generated many SARS-CoV-2 genome variants and which may affect the correct Reverse transcriptase-Real time Polymerase Chain Reaction (RT-PCR) diagnosis. Aim: To confirm and study the incidence of the non amplification of the SARS-CoV-2 Nucleocapsid gene (N gene) target among known SARS-CoV-2 positive samples. Materials and Methods: This retrospective observational study was carried out at the State Virology Laboratory, Gandhi Medical College, Bhopal, Madhya Pradesh, India during January 2021 to May 2021. During the study period, a total of 159 SARS-CoV-2 positive samples were failed to amplify the N gene target. To investigate the non amplification of N gene target of SARSCoV-2, a total of 20 samples were selected and retested using the initially used RT-PCR kit (VIRALDTECT RT-PCR kit) and also with the two different RT-PCR kits (TaqPath RT-PCR kit and Hi-PCR RT-PCR kit) which also contain primers/probes for the SARS-CoV-2 N gene target. Results: Amplification and detection of the SARS-CoV-2 N target gene was not observed in VIRALDTECT RT-PCR test results. In contrast, amplification was detected in the N gene target of SARS-CoV-2 while using the TaqPath and Hi-PCR kits. Obtained results confirm the failure of the annealing of VIRALDTECT kit N gene primer/probe and suggest the possible mutation event in the SARS-CoV-2 N gene among the N gene non amplified samples. Conclusion: Present study reports, the incidence of non amplification of SARS-CoV-2 N gene, where the RT-PCR kit failed to detect N gene target and seriously affect RT-PCR diagnosis. Hence, the study emphasises the revalidation of commercially available SARS-CoV-2, RT-PCR kits to identify these kinds of failure incidence.