ABSTRACT
Background: Serum interleukin 6 (IL-6) levels have been studied in the diagnostic evaluation of patients with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) disease (COVID-19). Methods: We studied the utility of treatment with tocilizumab in COVID-19 patients (n=19) with a negative nasopharyngeal swab real time reverse transcriptase polymerase chain reaction (RT-PCR) test for SARS-CoV-2 who had suggestive computed tomography (CT) findings, namely, COVID-19 Reporting and Data System (CO-RADS) 4,5. Results: Receiver operator characteristic (ROC) curve analysis showed that serum IL 6 at a cut-off of >56.9 pg/L was a predictor of mortality in nasopharyngeal swab RT-PCR negative patients with suggestive CT findings. Tocilizumab had no significant effect on the mortality. Conclusions: In nasopharyngeal swab RT-PCR negative patients with suggestive chest CT findings, elevated serum IL-6 levels > 56.9 pg/L predicted mortality. However, treatment with tocilizumab had no effect on mortality.
ABSTRACT
Background: The pandemic of COVID-19 since its beginning has created havoc all-round the globe. The role of oxygen therapy remains constant. Various modalities have been studied for oxygen delivery to hypoxic patients but high flow nasal oxygen (HFNO) has lately gained importance in terms of non-invasive oxygen delivery, easy administration and great improvement in patient's recovery. We conducted this retrospective analysis with the primary objective of looking for the proportion of patients who were successfully weaned off of HFNO or non-invasive ventilation (NIV) and the secondary aim was to look for duration of hospital stay and its effect on clinical recovery based on laboratory parameters. Methods: All patients, positive for COVID-19 infection by real-time reverse transcriptase polymerase chain reaction (RTPCR) were admitted to covid ICU or ward with oxygen requirement and were treated with either NIV or HFNO were enrolled for the study. Patients were grouped under H group (HFNO) or N group (NIV). Daily ABG readings, chest x-ray, respiratory rate, hemodynamic parameters and urine output were noted on 12 hourly intervals. Any changes in above parameters along with need for intubation were assessed. Results: Patients from both the groups showed significant improvement in their oxygen saturation by the fifth day of their treatment. Fourteen patients from the NIV group and 10 from the HFNO group had saturation >90% by Day 5. Of those who presented with saturation of <85%, 2 out of 5 in the NIV group (40%) and 1 of the 2 patients in the HFNO group (50%) showed improvement in their oxygen saturation. The P/F was statistically comparable (p 0.928) in both groups. The levels of bio markers, and the improvement was comparable and correlated with clinical improvement as well. Conclusion: We conclude that though HFNO is accepted better than NIV, the improvement in the respiratory status of the patient was comparable with both the treatment modalities and hence we do not recommend use of HFNO, especially in a situation of gross deficit of oxygen availability as compared to the exponential rise in the demand. © 2023 Tehran University of Medical Sciences.
ABSTRACT
To develop a multiplex fluorescent quantitative RT-PCR for the detection of porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV) and swine acute diarrhea syndrome coronavirus (SADS-CoV), in this study, specific primers/probes were designed based on the conserved regions of M, M and N gene sequences of PEDV, PDCoV and SADS-CoV, respectively. After optimization of the reaction conditions, a multiplex fluorescent quantitative RT-PCR for PEDV, PDCoV and SADS-CoV was established. The results of specificity assay showed that the method was positive for detection of PEDV, PDCoV and SADS-CoV, and negative for detection of porcine transmissible gastroenteritis virus, porcine rotavirus, porcine reproductive and respiratory syndrome virus, porcine pseudorabies virus, porcine circovirus type 2, porcine parvovirus, classical swine fever virus and foot-and-mouth disease virus. The results of sensitivity assay showed that the detection limit of this method for PEDV, PDCoV, and SADS-CoV plasmids standard was 1.0x101 copies/L, and had a good linear relationship with their Ct values in the range of 101 copies/L to 106 copies/L. The results of repeatability assay showed that the coefficients of variation (CVs) of intra- and inter-assay reproducibility ranged from 0.33% to 2.53%, indicating good repeatability and stability. To evaluate the effects of the developed method, 100 clinical samples collected from different parts of Henan province were used for detection of these three viruses and compared with those of single RT-PCR and standard methods. The results of multiplex fluorescent quantitative RT-PCR showed that the positive rates of PEDV, PDCoV and SADS-CoV were 38% (38/100), 14% (14/100) and 5% (5/100), respectively. There was no mixed infection. The coincidence rate with the standard detection methods of PEDV and PDCoV was 100%, and the sensitivity was higher than that of single RT-PCR. In this study, a specific, sensitive and rapid multiplex fluorescent quantitative RTPCR method was established for the first time, which could be used for the differential detection of PEDV, PDCoV and SADS-CoV, and laid a foundation for the differential diagnosis and control of porcine diarrheal diseases.
ABSTRACT
Purpose: Compared to nasopharyngeal/oropharyngeal swabs (N/OPS-VTM), non-invasive saliva samples have enormous potential for scalability and routine population screening of SARS-CoV-2. In this study, we investigate the efficacy of saliva samples relative to N/OPS-VTM for use as a direct source for RT-PCR based SARS-CoV-2 detection. Method(s): We collected paired nasopharyngeal/oropharyngeal swabs and saliva samples from suspected positive SARS-CoV-2 patients and tested using RT-PCR. We used generalized linear models to investigate factors that explain result agreement. Further, we used simulations to evaluate the effectiveness of saliva-based screening in restricting the spread of infection in a large campus such as an educational institution. Result(s): We observed a 75.4% agreement between saliva and N/OPS-VTM, that increased drastically to 83% in samples stored for less than three days. Such samples processed within two days of collection showed 74.5% test sensitivity. Our simulations suggest that a test with 75% sensitivity, but high daily capacity can be very effective in limiting the size of infection clusters in a workspace. Guided by these results, we successfully implemented a saliva-based screening in the Bangalore Life Sciences Cluster (BLiSC) campus. Conclusion(s): These results suggest that saliva may be a viable alternate source for SARS-CoV-2 surveillance if samples are processed immediately. Although saliva shows slightly lower sensitivity levels when compared to N/OPS-VTM, saliva collection is logistically advantageous. We strongly recommend the implementation of saliva-based screening strategies for large workplaces and in schools, as well as for population-level screening and routine surveillance as we learn to live with the SARS-CoV-2 virus.Copyright © 2023 Indian Association of Medical Microbiologists
ABSTRACT
Background and Aim: Feline infectious peritonitis (FIP) is an infectious, immune-mediated, and fatal disease in cats caused by a mutant feline coronavirus (FCoV) infection. Feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) are two common retroviruses that play a role in reducing feline immune function with opportunistic retrovirus infection being a predisposing factor for the development of FIP. This study aimed to evaluate the clinicopathological parameters of FIP in cats with and without retrovirus coinfection. Materials and Methods: In total, 62 cats presenting with pleural and/or peritoneal effusion at the Kasetsart University Veterinary Teaching Hospital, Bangkok, Thailand, were selected for the study. Effusion samples were collected and a reverse transcriptase-polymerase chain reaction (RT-PCR) assay was performed on all samples using the 3' untranslated region primer. All FCoV-positive cats were tested for retrovirus infection using a commercial kit (Witness FeLV-FIV [Zoetis]; United States). Clinical signs, hematological, and biochemical parameters of these cats were investigated and grouped. Results: Of the 62 cats with pleural and/or peritoneal effusion, FCoV was detected in 32, of which 21 were highly suspicious for FIP. The cats suspected of FIP were divided into three subgroups following viral detection. A total of 14 had only FCoV infection (Group A), four had FCoV and FeLV infection (Group B), and three had FCoV, FeLV, and FIV infection (Group C). Of the rest, 11 had definitive diagnoses, which included three being FCoV and FeLV-positive (Group D), and eight were retrovirus-negative (Group E). Mild anemia and lymphopenia were found in cats infected with these three viruses. An albumin-to-globulin ratio lower than 0.5 was found in FIP cats with only FCoV infection. Conclusion: Typically, cats with clinical effusion and FIP, with and without retrovirus coinfection, had similar hematological findings. Clinical signs, blood parameters, fluid analysis with cytological assessment, and RT-PCR assays could identify better criteria to diagnose FIP with and without retrovirus coinfection.
ABSTRACT
Given the scale of the ongoing COVID-19 pandemic, the need for reliable, scalable testing, and the likelihood of reagent shortages, especially in resource-poor settings, we have developed an RTqPCR assay that relies on an alternative to conventional viral reverse transcriptases, a thermostable reverse transcriptase/DNA polymerase (RTX) (Ellefson et al., 2016). Here we show that RTX performs comparably to the other assays sanctioned by the CDC and validated in kit format. We demonstrate two modes of RTX use - (i) dye-based RT-qPCR assays that require only RTX polymerase, and (ii) TaqMan RT-qPCR assays that use a combination of RTX and Taq DNA polymerases (as the RTX exonuclease does not degrade a TaqMan probe). We also provide straightforward recipes for the purification of this alternative reagent RTX. We anticipate that in low resource or point-of-need settings researchers could obtain the available constructs and begin to develop their own assays, within whatever regulatory framework exists for them.Copyright © 2021 Bio-protocol LLC. All Rights Reserved.
ABSTRACT
Bovine rhinitis virus (BRV) is an important pathogen responsible for the bovine respiratory disease complex (BRDC) and can be divided into two genotypes (BRAV and BRBV). To establish a duplex quantitative real-time RT-PCR assay for simultaneous detection of BRAV and BRBV, specific primers and TaqMan probes targeting the 5'NTR of BRAV and 3'NTR of BRBV were designed. A duplex quantitative real- time RT- PCR assay for simultaneous detecting BRAV and BRBV was preliminarily established by optimizing reaction conditions for each step. The assay specifically detects BRAV and BRBV, and no crossreaction with other common bovine respiratory pathogens, including IDV, BCoV, BVDV-1, BRSV, BPIV-3, BAdV-3, mycoplasma bovis, Pasteurella multocida, Mannheimia haemolytica, Escherichia coli, and Salmonella, was observed. In addition, the sensitivity test showed that the detection limits of this assay were 3.2x101 copies/L for both BRAV and BRBV plasmid standards. Besides, the repeatability test showed that the variation coefficients of this assay were less than 0.05 from both lot-to-lot and intra-lot. These results showed that the assay has high specificity, extreme sensitivity, and good repeatability. Moreover, a total of 43 nasal swabs of BRDC cattle were tested by our assay and four other quantitative real-time RT-PCR assays, including 3 BRAV assays and 4 BRBV assays. The results showed that the detection rates of our assay were 32.56%(14/43) for BRAV and 30.23%(13/43) for BRBV, and the detection rates of other quantitative real-time RT-PCR assays were 0(0/43), 2.33%(1/43), 23.26%(10/43) for BRAV and 27.91% (12/43), 27.91%(12/43), 27.91%(12/43), 27.91%(12/43) for BRBV, indicating that our assay has a more substantial detection capability than other assays. This study firstly established a duplex quantitative real-time RT-PCR assay for simultaneous detection of BRAV and BRBV, and the assay exhibited high specificity, sensitivity, and stability. Moreover, the study firstly confirmed the existence of BRAV in China, contributing to the prevention and control of BRDC.
ABSTRACT
The emergence of SARS-CoV-2 necessitated the rapid deployment of tests to diagnose COVID-19. To monitor the accuracy of testing across the COVID-19 laboratory network in Thailand, the Department of Medical Sciences under the Ministry of Public Health launched a national external quality assessment (EQA) scheme using samples containing inactivated SARS-CoV-2 culture supernatant from a predominant strain in the early phase of the Thailand outbreak. All 197 laboratories in the network participated; 93% (n=183) of which reported correct results for all 6 EQA samples. Ten laboratories reported false-negative results, mostly for samples with low viral concentrations, and 5 laboratories reported false-positive results (1 laboratory reported false positives and false negatives). An intralaboratory investigation of 14 laboratories reporting incorrect results revealed 2 main causes of error: (1) RNA contamination of the rRT-PCR reaction and (2) poor-quality RNA extraction. Specific reagent combinations were significantly associated with false-negative reports. Thailand's approach to national EQA for SARS-CoV-2 can serve as a roadmap for other countries interested in implementing a national EQA program to ensure laboratories provide accurate testing results, which is crucial in diagnosis, prevention, and control strategies. A national EQA program can be less costly and thus more sustainable than commercial EQA programs. National EQA is recommended to detect and correct testing errors and provide postmarket surveillance for diagnostic test performance.
Subject(s)
COVID-19 , Humans , SARS-CoV-2/genetics , Laboratories , Pandemics/prevention & control , Thailand/epidemiology , RNA, Viral/geneticsABSTRACT
Background: The recent transmission clusters (RTCs) identified through phylogenetic approaches allow to describe the main transmission networks. This render possible to describe potential shifts among HIV transmission routes and populations and, in some cases, to specifically target prevention measures. Here we describe the evolution of RTCs over the last decade in a specialized laboratory serving centers from the entire French territory. Method(s): We extracted all the HIV reverse transcriptase sequences available between 01/01/2013 and 31/08/2022. The sequences dataset was studied overall and divided into three equal time periods: 2013-15, 2016-18, 2019-2021. The first sequences available for each patient were aligned and the trees were reconstructed by maximum likelihood using IQtree software. Clusters, defined by a maximum genetic distance < 4.5% and a branch support >90%, were extracted using ClusterPicker. Result(s): Overall, 8591 sequences were included. Among them, 950 RTCs were identified including 2492 sequences (29%) and 68 large RTCs ( >4 sequences) with 475 (5.6%) sequences. The mean duration of large RTCs (from the first to the last sequences) was 5.1 years [IQR: 4.1-7.1] and 34 were still active (including at least one sequence during the last year of the study period). 3640, 2897 and 2157 sequences were included for the 2013-15, 2016-18 and 2019-2021 periods, respectively. We identified 298 RTCs (19.5% of sequences), 249 (20.4%) and 226 (27.5%) among those periods, respectively. While the number of sequence pairs decreased from 2013-15 to 2019-21, the number of large RTCs increased steadily (see Table 1). During the period 2019-21, including the largest clusters, patients belonging to a RTC were more often male (68 vs 58%, p< 0.001) and younger (average age: 39 vs 44 years, p< 0.001) than non-RTC patients. This observation was even more marked for very large RTCs (see Table 2). It should be noted that the largest cluster (14 patients) was mainly composed of women and located in French overseas territories. Conclusion(s): This study shows an evolution of the structure of HIV sequence clusters over time with a decreasing number of small RTCs but an increasing number of large RTCs. These trends can be explained by a better global control of transmission, due in part to TasP, but not preventing some super-transmitters networks, despite PrEP use and not only including MSM is some settings. The COVID period does not seem to have strongly prevented such large transmission networks.
ABSTRACT
Objective: Epidemiology and genetic variations of the infectious bronchitis virus(IBV) in Fujian province were studied. Method: Two strains of virus isolated from the diseased chickens in Fujian in 2021 were identified by chicken embryo pathogenicity test, electron microscope observation, and RT-PCR. S1 genes of the isolates were cloned, sequenced, and analyzed using biological software. Result: The two IBV strains were code named FJ-NP01 and FJ-FZ01. The full length of S1 of FJ-NP01 was 1 629 nt encoding 543 amino acids, and that of FJ-FZ01, 1 620 nt encoding 540 amino acids. The S1 gene cleavage site of FJ-FZ01 was HRRRR, same as all reference strains of genotype I branch;while that of FJ-NP01 HRRKR differed from the reported site of IBV isolated from genotype IV but same as that of TC07-2 reference strain of genotype VI. The homology of nucleotide and amino acid between the two isolates was 83.2% and 79.6%, respectively, but merely 75.7%-76.3%and 77.1%-83.5% with the Mass-type conventional vaccines H120 and H52, respectively. Further analysis showed that FJ-NP01was from a recombination event between CK CH GD LZ12-4 and L-1148, the homology of nucleotide acid between 1438-1506 nt of FJ-NP01 with CK CH GD LZ12-4 was 97%, and 95.9% between the other nucleotide acid of S1 gene with L-1148. Conclusion: It appeared that the IBV epidemic experienced in the province was complex in nature and that the existing Mass vaccines would not provide sufficient immune protection to deter the spread.
ABSTRACT
In order to perform the isolation of avian infectious bronchitis virus (IBV) and study the pathogenicity of IBV isolate, the RT-PCR was used to detect nucleic acid extracted from a clinical sample of chickens, which were suspected to be infected with infectious bronchitis virus (IBV) and provided by a farmer in Yuncheng, Shanxi province. And the sample was detected as IBV positive by RT-PCR. Then 9-11-day-old SPF chicken embryonated eggs were inoculated with the sample filtered from the grinding fluid, and the obtained allantoic fluid was blindly passed by three generations (F3) and was also tested as IBV positive;The F11 generation passaged in embryonated eggs caused typical "dwarf embryo" lesions to SPF chicken embryonated eggs, and induced the loss of cilia in tracheal rings. The results showed that an IBV strain was isolated and named as YC181031. The S1 gene amplification and sequencing analysis showed that YC181031 strain belonged to IBV GI-22 genotype, which is also nephropathogenic type IBV. Seven-day-old SPF chicks were used to test the pathogenicity of the isolate. The results showed that several clinical symptoms were showed in chicks infected with YC181031, such as breathing with difficulty, depression, excreting watery droppings and death. The mortality of infected chicks was 20%. Typical pathological changes such as enlargement of kidney and urate deposition in the kidney were observed in infected chicks. The immunohistochemical assay and viral load detection were performed for the tissue samples from infected and dead chicks. The tissue lesions and distribution of virus were observed in the kidney, trachea, lung, glandular stomach, spleen and liver samples of infected chicks. RT-PCR detection of pharyngeal anal swabs showed that the virus shedding by infected chicks could be continuously detected within 14 days of the test period;The viral loads of various tissues were detected by RT-qPCR and the results showed that the viral load from high to low was kidney, trachea, lung, stomach, spleen and liver. The viral load of kidney was significantly higher than that of other tissues (P < 0.05).In this study, the pathogenicity characteristics of GI-22 genotype strain were systematically studied for the first time, providing a reference for the prevention and treatment of the disease.
ABSTRACT
Background: The COVID-19 pandemic has led to a dramatic loss of human life worldwide and presents an unprecedented challenge for healthcare systems worldwide. Earlier to SARS-CoV pandemic, coronaviruses were only thought to cause mild, self-limiting upper respiratory tract infections in humans. COVID 19 presents across a spectrum of symptoms. WHO recommends detection of unique sequences of virus RNA by Nucleic Acid Amplification Test (NAAT) such as real-time reverse-transcriptase polymerase chain reaction (rRT-PCR). The aim of this cross sectional study was analysis and confirmation of Nasopharyngeal/oropharyngeal swab specimen by real-time reverse transcription polymerase chain reaction (RT-PCR). Material & Methods: This was a cross-sectional retrospective study that reviewed records of samples collected from June 2021 to March 2022. Nasopharyngeal/oropharyngeal swab specimen were collected from suspected COVID-19 subjects of various districts of Punjab and referred to Viral Research Diagnostic Laboratory [VRDL], Government Medical College [GMC], Amritsar for laboratory analysis and confirmation by real-time reverse transcription polymerase chain reaction (RT-PCR). Results: During the present study, a total of 11,27,005 samples were analyzed from June 2021 to March 2022 for SARS-CoV-2 detection by ICMR approved COVID-19 RT-PCR kits. Out of total 11,27,005 cases, 24,466 cases (2.17%) were found to be SARS-CoV-2 positive while 11,02,539 cases (97.83%) were SARS-CoV-2 negative. Conclusions: Ever since the COVID-19 global pandemic emerged, the developing countries are facing challenges regarding its diagnosis. Isolation of the infected person will eventually decrease the Reproduction number i.e Ro which will further interrupt the transmission cycle leading to decrease in community spread.
ABSTRACT
Background: Coronavirus disease 2019 (COVID-19) was a recent global pandemic of the era which posed a great challenge for the health care in terms of preventive, diagnostic and treatment dimensions. The seroprevalence rate of COVID IgG antibodies is very crucial in estimating the susceptibility of a particular area to the viral disease. In our study, we estimated the seroprevalence of COVID-19 in a rural area. Aims and Objectives: We aimed to estimate the seroprevalence of COVID-19 in a rural district of Tamil Nadu, 6 months after the index case. Materials and Methods: We conducted a cross-sectional study of 509 adults aged more than 18 years. From all the seven Taluks, two gram panchayats (administrative cluster of 8-10 villages) were randomly selected followed by one village through convenience. The participants were invited for the study to the community-based study kiosk set up in all the eight villages through village health committees. We collected sociodemographic characteristics and symptoms using a mobile application-based questionnaire, and we tested samples for the presence of IgG antibodies for severe acute respiratory syndrome coronavirus 2 using an electro chemiluminescent immunoassay. We calculated age-gender adjusted and test performance adjusted seroprevalence. Results: The age-and gender-adjusted seroprevalence was 8.5% (95% confidence interval [CI] 6.9-10.8%). The unadjusted seroprevalence among participants with hypertension and diabetes was 16.3% (95% CI: 9.2-25.8) and 10.7% (95% CI: 5.5-18.3), respectively. When we adjusted for the test performance, the seroprevalence was 6.1% (95% CI 4.02-8.17). The study estimated 7 (95% CI 1:4.5-1:9) undetected infected individuals for every reverse transcription polymerase chain reaction confirmed case. Infection fatality rate (IFR) was calculated as 12.38/10,000 infections as on October 22, 2020. History of self-reported symptoms and education were significantly associated with positive status (P<0.05). Conclusion: A significant proportion of the rural population in a district of Tamil Nadu remains susceptible to COVID-19. A higher proportion of susceptible, relatively higher IFR, and a poor tertiary health-care network stress the importance of sustaining the public health measures and promoting early access to the vaccine are crucial to preserving the health of this population. Low population density, good housing, adequate ventilation, limited urbanization combined with public, private, and local health leadership are critical components of curbing future respiratory pandemics. [ FROM AUTHOR] Copyright of Asian Journal of Medical Sciences is the property of Manipal Colleges of Medical Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full . (Copyright applies to all s.)
ABSTRACT
Shortly after its emergence, Omicron and its sub-variants have quickly replaced the Delta variant during the current COVID-19 outbreaks in Vietnam and around the world. To enable the rapid and timely detection of existing and future variants for epidemiological surveillance and diagnostic applications, a robust, economical real-time PCR method that can specifically and sensitively detect and identify multiple different circulating variants is needed. The principle of target- failure (TF) real-time PCR is simple. If a target contains a deletion mutation, then there is a mismatch with the primer or probe, and the real-time PCR will fail to amplify the target. In this study, we designed and evaluated a novel multiplex RT real-time PCR (MPL RT-rPCR) based on the principle of target failure to detect and identify different variants of SARS-CoV-2 directly from the nasopharyngeal swabs collected from COVID-19 suspected cases. The primers and probes were designed based on the specific deletion mutations of current circulating variants. To evaluate the results from the MPL RT-rPCR, this study also designed nine pairs of primers for amplifying and sequencing of nine fragments from the S gene containing mutations of known variants. We demonstrated that (i) our MPL RT-rPCR was able to accurately detect multiple variants that existed in a single sample; (ii) the limit of detection of the MPL RT-rPCR in the detection of the variants ranged from 1 to 10 copies for Omicron BA.2 and BA.5, and from 10 to 100 copies for Delta, Omicron BA.1, recombination of BA.1 and BA.2, and BA.4; (iii) between January and September 2022, Omicron BA.1 emerged and co-existed with the Delta variant during the early period, both of which were rapidly replaced by Omicron BA.2, and this was followed by Omicron BA.5 as the dominant variant toward the later period. Our results showed that SARS-CoV-2 variants rapidly evolved within a short period of time, proving the importance of a robust, economical, and easy-to-access method not just for epidemiological surveillance but also for diagnoses around the world where SARS-CoV-2 variants remain the WHO's highest health concern. Our highly sensitive and specific MPL RT-rPCR is considered suitable for further implementation in many laboratories, especially in developing countries.
ABSTRACT
OBJECTIVE: To investigate and analyze multiple detection of 13 kinds of viruses in 500 children with acute respiratory tract infection in Hami of Xinjiang. METHODS: A total of 500 children with acute respiratory tract infection treated in the hospital between Jan 2018 and Jan 2021 were enrolled. Thirteen kinds of respiratory infection viruses including human respiratory syncytial virus(RSV), human rhinovirus(hRV), respiratory adenovirus(AdV), influenza A and B viruses(Inf A, Inf B), parainfluenza virus(PIV 1/2/3), human enterovirus(hEV), human metapneumovirus(hMPV), human coronavirus(hCoV 229E/OC43) and human Boca virus(hBoV) were detected by multiple reverse transcription polymerase chain reaction(RT-PCR) amplification and capillary electrophoresis. And the results were compared with those by direct sequencing method. RESULTS: Of the 500 samples, 379 samples were positive(75.80%), and the top three detection rates were RSV(19.40%), hRV(16.00%) and Inf B(12.60%). The differences in positive rates of the respiratory virus among <1 year group, 1-3 years group and >3 years group were significant(84.97%, 77.47%, 65.45%)(P<0.05). The detection rate of RSV was the highest in <1 year group, and the detection rates of Inf A and Inf B were the highest in >3 years group. The differences in positive rates of respiratory viruses among the spring group, summer group, autumn group and winter group were significant(74.05%, 63.73%, 77.24%, 84.03%)(P<0.05). The detection rates of RSV, PIV 3, and hMPV were the highest in the winter group, and detection rate of AdV was the highest in spring group. CONCLUSION: RSV is the main infection virus in children with acute respiratory infection in Hami of Xinjiang. The distribution of respiratory viruses is related to age and onset season in children.
ABSTRACT
Infectious peritonitis virus (FIPV) causes a fatal disease in cats. This virus occurs both in cats bred in households with optimal welfare and outdoor cats. Feline patients with the effusive form of disease usually survive a few days to weeks from the appearance of the first clinical signs. Cats with the non- effusive form survive for weeks to months. FIPV is caused by a mutation from feline enteric coronavirus (FECV). In our study, we diagnosed feline coronavirus from the feces of 82% of the tested cats. The persistence of the feline coronavirus in the organism is influenced by environmental factors, the genome of the host and the causative agent. Negative environmental conditions that increase the likelihood of FIPV disease are long-term stress, mainly more labile individuals and a high concentration of domesticated cats in one place. In the host, there are important factors such as immune system performance, age, breed and genetic background. In our study, we primarily verified the real time RT-PCR method for identifying the virus from the feces of 71 cats and subsequently gaine the valuable data on the dynamics of feline coronavirus excretion, primarily for epizootological purposes and for the purposes of genetic analyzes of susceptibility to infection.
ABSTRACT
Background: To assess role of HRCT, RT-PCR in the evaluation of COVID-19 symptomatic patients. Material and Methods: This retrospective study was conducted in the department of Radiology and Microbiology at Aarupadaiveedu Meddical College and Hospital . Eighty- five clinically suspects of COVID-19 patients of either gender was enrolled in the study and underwent RT-PCR test and HRCT Thorax . HRCT images were studied by Three radiologists independently. CURB- 65 and SOFA was recorded. Results: There were 50 males and 35 females in our study. Disease severity CURB was 1 and SOFA was 3. Chest CT had suspicious for COVID-19 in 45 and RT-PCR SARS-CoV2 positive in 40.42 had no comorbidity, 8 had peripheral vascular disease, 12 had cerebrovascular disease 13 had hypertension and 10 had diabetes mellitus. The difference was significant (P< 0.05). In all 40 patients of PCR SARS-CoV-2 positive, 35 HRCT were judged as suspicious for COVID-19. In 45 cases of PCR SARS-CoV-2 negative, 10 HRCT scans were judged as not suspicious for COVID-19. HRCT Thorax had a sensitivity of 87.5%, specificity of 77.8%, PPV of 77% and NPV of 87.5%. Conclusion: The diagnostic accuracy of HRCT Thorax in symptomatic patients is good, but not good enough to safely diagnose or exclude COVID-19 patients. RT- PCR is useful in addition with HRCT in diagnosis of COVID-19 patients.
ABSTRACT
It has been proven that mRNA vaccines are highly effective against the COVID-19 outbreak, and low prevalence of side effects has been shown. However, there are still many gaps in our understanding of the biology and biosafety of nucleic acids as components of lipid nanoparticles (LNPs) most often used as a system for inctracellular delivery of mRNA-based vaccines. It is known that LNPs cause severe injection site inflammation, have broad biodistribution profiles, and are found in multiple tissues of the body, including the brain, after administration. The role of new medications with such pharmacokinetics in inflammation developing in inaccessible organs is poorly understood. The study was aimed to assess the effects of various doses of mRNA-LNP expressing the reporter protein (0, 5, 10, and 20 microg of mRNA encoding the firefly luciferase) on the expression of neuroinflammation markers (Tnfalpha, Il1beta, Gfap, Aif1) in the prefrontal cortex and hypothalamus of laboratory animals 4, 8, and 30 h after the intramuscular injection of LNP nanoemulsion. It was shown that mRNA-LNP vaccines in a dose of 10-20 microg of mRNA could enhance Aif1 expression in the hypothalamus 8 h after vaccination, however, no such differences were observed after 30 h. It was found that the Gfap, l11beta, Tnfalpha expression levels in the hypothalamus observed at different times in the experimental groups were different. According to the results, mRNA-LNPs administered by the parenteral route can stimulate temporary activation of microglia in certain time intervals in the dose-dependent and site specific manner.Copyright © 2022 Pirogov Russian National Research Medical University. All rights reserved.
ABSTRACT
Aim: COVID-19 infection has affected the whole world. It has been speculated that the virus might hold on to angiotensin-converting enzyme 2 (ACE 2) surfaces of type 2 alveolar cells. ACE inhibitors and angiotensin receptor antagonists (ARBs) are essential antihypertensive and cardiac failure drugs in the guidelines. In this study, we aimed to find the effect of these drugs on clinical, laboratory courses, and outcomes of COVID-19 patients. Material(s) and Method(s): We included 109 patients in this study. There were 43 patients in the ACE/ARB group and 66 patients in the non-ACE/ARB group. The mean age was 60 years in the ACE/ARB group and 52 years old in the non-ACE/ARB group. Basal symptoms, hemogram, CRP, D-dimer, LDH, Ferritin, AST, duration of hospitalization, percentage of intensive care unit (ICU) need, length of stay in ICU were compared between the groups. Result(s): The mean age in the ACE/ARB group was higher than in the other group and was statistically significant (p=.027). The initial symptoms were not different. There were no differences between the laboratory results of the groups. The ICU need was higher in the patients who do not use the drug than in the users (p<.020). Discussion(s): ACE/ARB usage in COVID-19 patients did not worsen the course of the disease. However, ACE/ARB users before COVID-19 pandemic were taken to ICU at a low rate.Copyright © 2022, Derman Medical Publishing. All rights reserved.
ABSTRACT
Although real‐time quantitative reverse transcription polymerase chain reaction (RT‐qPCR) is the gold standard for detecting the virus severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) and other pathogens, the coronavirus disease 2019 (COVID‐19) pandemic has highlighted the scarcity of instruments, devices, and reagents for polymerase chain reaction (PCR) testing in constrained settings. At least for under‐resourced countries, it has become critical to deploy instruments that can be rapidly constructed and satisfy this demand. Instead of separating the optical system from the thermal module (typical of qPCR thermocyclers), we report a portable Hybrid Opto‐Thermocycler—dubbed HybOT Cycler—that takes advantage of the high‐temperature tolerances (>100 °C) of electronic and optical components to combine thermal control, illumination, and fluorescence detection into a highly integrated hybrid module. This simple configuration allowed us to reduce the overall number of components, thus simplifying its assembly and reducing the instrument size. The HybOT Cycler is wirelessly controlled from an application installed in a tablet. PCR assays are carried out in a bubble‐free microfluidic device that can be easily replicated from an acrylic mold. Using the HybOT Cycler, down to 100 copies/µL of genetic material of the virus SARS‐CoV‐2 with 95% sensitivity and 100% specificity is detected. The HybOT Cycler can assist in diagnosing SARS‐CoV‐2 and other pathogens in resource‐poor settings. [ FROM AUTHOR] Copyright of Advanced Materials Technologies is the property of John Wiley & Sons, Inc. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full . (Copyright applies to all s.)