Real-time RT-PCR Test Research Articles

Global distribution of sporadic sapovirus infections: A systematic review and meta-analysis.

Acute gastroenteritis (AGE), characterized by diarrhea and vomiting, is an important cause of global mortality, accounting for 9% of all deaths in children under five years of age. Since the reduction of rotavirus in countries that have included rotavirus vaccines in their national immunization programs, other viruses such as norovirus and sapovirus have emerged as more common causes of AGE. Due to widespread use of real-time RT-PCR testing, sapovirus has been increasingly reported as the etiologic agent in both AGE outbreaks and sporadic AGE cases. We aimed to assess the role of sapovirus as a cause of endemic AGE worldwide by conducting a systematic review of published studies that used molecular diagnostics to assess the prevalence of sapovirus among individuals with AGE symptoms. Of 106 articles included, the pooled sapovirus prevalence was 3.4%, with highest prevalence among children <5 years of age (4.4%) and among individuals in community settings (7.1%). Compared to studies that used conventional RT-PCR, RT-qPCR assays had a higher pooled prevalence (5.6%). Among individuals without AGE symptoms, the pooled sapovirus prevalence was 2.7%. These results highlight the relative contribution of sapovirus to cases of AGE, especially in community settings and among children <5 years of age.

In vitro Inhibition of Border Disease Virus Replication With Lentivirus-Mediated shRNAs.

Background: Border disease is believed to be one of the most important diseases in the animal husbandry industry, which has not yet been eradicated in Iran. The development of approaches based on the application of interfering RNA (RNAi) for antiviral therapy has attracted a great deal of attention over the recent years. The present research was conducted to design, construct, and apply shRNA against the NS3 gene of BDV to evaluate the prevention of BDV proliferation in the cell culture system. For this purpose, the suitable oligonucleotide sequence of NS3 gene coding was selected utilizing BDV- X818 strain. Afterwards, using shRNA design software, shRNA molecules were designed and synthesized. These shRNAs were cloned into the desired vectors and were finally transfected in HEK293T cells employing the third generation of lentiviral packaging system. Subsequently, these shRNA expressing lentiviruses were transduced to the MDBK cell line to challenge to border virus. In order to evaluate the efficacy of shRNAs, the viral infectious titer and RNA copy number were calculated with TCID50 and Real-time RT-PCR tests, respectively.Results: The results revealed that shRNAs 1, 2, and 3 decreased viral RNA by more than 90% compared to the control groups. BDV titer noticeably decreased after the challenge with shRNAs 1, 2, and 3 from ~88% up to 99% in comparison with the control groups.Conclusions: Overall, it could be concluded that RNAi may be considered as a strong treatment proposal against viruses, such as BDV.

Evaluation of SARS-CoV-2 RT-PCR test results from a pandemic hospital according to demographic data

ObjectivesThe emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) leading to coronavirus disease 2019 (COVID-19) in China at the end of 2019 has resulted in a global pandemic. On 11 March 2020, the first case of COVID-19 was reported in Turkey. The aim of this study was to evaluate SARS-CoV-2 Real-Time Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) test results from the Medical Microbiology Laboratory of a pandemic hospital according to demographic data. Study designRetrospective cohort study. MethodsSARS-CoV-2 RT-PCR test results of 413,013 samples from 194,062 patients were retrospectively analysed. Tests were carried out between 27 March and 31 December 2020 using two commercial kits. The patient's age and gender were recorded, in addition to the percentage of positive test results per month (i.e. monthly positivity). Pearson’s Chi-squared test was used to analyse statistical significance. ResultsOverall SARS-CoV-2 positivity in the pandemic hospital was 19.9%. Female gender and younger age (0–18 years) had a statistically significant higher positivity (P < 0.05). There was a statistically significant higher positivity in August and September. ConclusionsHigher positivity among the younger population and females may be the leading cause of low COVID-19 mortality rates in Turkey as these population groups are less likely to die from the disease. Governments should disaggregate COVID-19 data by age and gender, and vaccine studies focussing on younger populations should be accelerated because this population group represents an important source of infection.

COVID-19 Pandemic: A comparison of adult and pediatric populations

Aim: Couple of pneumonia cases were reported in a short period in Wuhan, China. The cases were revealed to be associated with a different coronavirus type was named SARS-CoV-2 and the disease was identified as Covid-19. It is known that the disease occurs in all age groups. We aimed to evaluate the differences in clinical and laboratory features between adult and pediatric patients. Method: The study is a retrospective cross-sectional study and consists of 206 patients with a definitive diagnosis of Covid-19 confirmed by a positive real-time reverse-transcriptase polymerase chain reaction (RT-PCR) testing for SARS-CoV-2. They hospitalized in health institutions connected to City Health Administrative of Bolu, Turkey between 11.03.2020 and 19.04.2020. The number of the pediatric patients (0-17 age group) was 106, that of the adult patients (18 and above age group) was 100. Data concerning the patients consisted of age, symptoms, laboratory parameters such as hemogram, biochemistry, coagulation. Results: The mean values of platelet (PLT), platocrit (PCT), percentage of monocytes (MONO %) and MONO in the “0-17” age group were found to be significantly higher than the mean values in the “18 and above” age group. The hemoglobin (HGB) mean value of the “0-17” age group was significantly lower than that of the “18 and above” age group. The mean C - reactive protein (CRP) value of the “0-17” age group (4.55; min: 1.20 – max: 11.80) was significantly lower than that of the “18 and above” age group (5.35; min 1.25- max: 19.77) with no statistically significance. In pediatric group, the most common symptoms were other symptoms like diarrhea, vomiting and joint pain whereas the adult patients had fever and cough often with statistically significant. Conclusion: Clinical findings and laboratory abnormalities in Covid-19 are less common in children. Although it seems that Covid-19 is less symptomatic in children, they are also affected by the disease. Performing RT-PCR test based on the contact history of the children may help to minimize morbidity with an early diagnosis. Multicenter studies with more numbers of patients should be performed.

ПРОТОКОЛ ДЕТЕКЦІЇ ГЕНЕТИЧНИХ МАРКЕРІВ У СЛИНІ, ВИКОРИСТОВУЮЧИ ПОЛІМЕРАЗНУ ЛАНЦЮГОВУ РЕАКЦІЮ БЕЗ ОЧИСТКИ НУКЛЕЇНОВИХ КИСЛОТ: ТЕСТУВАННЯ SARS-COV-2 ТА GAPDH МАРКЕРІВ

Introduction. Polymerase chain reaction (PCR)-based diagnostic tests use purifi ed nucleic acids (NAs) from clinical samples. The NAs purifi cation step adds time, cost, and aff ects the quality of testing. The objective of this study was to develop a protocol for direct use of saliva in tests for genetic markers, without purifi cation of nucleic acids. Methods. PCR, real-time RT-PCR and isothermal amplifi cation tests were used for direct detection of genetic markers, without purifi cation of nucleic acids. Results. We report a protocol for the direct detection of genetic markers in saliva. The protocol is based on a collection of saliva in a solution containing a detergent and ethanol and is compatible with isothermal amplifi cation (LAMP), real-time RT-PCR and RT-PCR. SARS-CoV-2 and GAPDH markers were used as reference markers. We observed that mild detergents allow effi cient detection of external reference and intracellular endogenous markers, while strong detergent, e.g. sodium dodecyl sulfate, inhibited the PCR reaction. Under these conditions, saliva samples can be stored for 24 h at +4°C or –18°C with the preservation of markers. Storage at room temperature led to the deterioration of marker detection. Snap heating of saliva samples at the time of collection, followed by storage at room temperature, provided partial protection. Conclusion. The protocol presented in this report describes the collection and storage of saliva for direct detection of genetic markers and is compatible with PCR and LAMP tests.

Sero-molecular epidemiology of hepatitis E virus in pigs and human contacts in Ghana

BackgroundHepatitis E virus (HEV) is among the leading causes of viral hepatitis in most developing countries. Zoonotic acquisition of HEV genotype 3 from swine has come into focus more recently. Available studies on HEV in Ghana and other countries in the region do not provide enough information towards understanding the epidemiology of HEV in human and animal populations. Towards this end, we conducted a comparative cross-sectional study to determine the seroprevalence and risk factors associated with HEV exposure, both in swine and humans working on pig farms in typical local settings. The presence of viral RNA in human and swine samples was also evaluated, along with classification of viral sequences from HEV-positive samples.MethodsStructured questionnaires soliciting information on pigs reared, as well as socio-demographic information including age, sex and educational background of humans was collected. A total of 10 ml and 5 ml of whole blood was collected from pigs and human participants respectively. ELISA and real-time RT-PCR were performed on the sera for the qualitative detection of IgG antibodies to hepatitis E virus and viral RNA, respectively.ResultsFive hundred and forty-four (544) human participants including 264 swine contacts and 280 swine non-contacts were enrolled in the study. Although the proportion of HEV IgG antibodies was higher in contact groups (114; 54.3%) than non-contact groups (96; 45.7%), a multivariate analysis did not show any significant difference. No HEV RNA was detected in human samples. Similarly, 720 pigs were sampled from 18 farms located in five regions in Ghana. Twenty-three (23) of the pigs (3.2, 95%CI = 2.0–4.8) were positive for HEV RNA by real-time RT-PCR testing. Sequences obtained from HEV-positive samples were found to share high sequence identities with each other and clustered with other genotype 3 viruses indicating the existence of circulating zoonotic genotype 3 viruses on farms. Although we did not find evidence of pig to human transmission of HEV genotype 3, the presence of this genotype in pigs shows the potential for possible zoonotic transmission in African farm settings and buttresses the importance of active surveillance for the infection among at risk populations.

Critical parameters of real time reverse transcription polymerase chain reaction (RT-PCR) diagnostics: Sensitivity and specificity for bluetongue virus

A new variant of bluetongue virus serotype 3, BTV3 ITL 2018 (here named: BTV3), was included in serial dilutions in the BT Proficiency Test 2020. Although the OIE-recommended panBTV real time RT-PCR test targeting genome segment 10 (Seg-10) detected this variant, we showed that reverse transcription (RT) at 61 °C instead of 50 °C completely abolished detection. Another Seg-10 panBTV real time RT-PCR test detected BTV3, irrespective of the temperature of RT. In silico validation showed that each of the OIE-recommended PCR primers using IVI-primers contain single mismatches at the -3 position for BTV3. In contrast, WBVR-primers of a second test completely match to the BTV3 variant. Our results suggest that single mismatches caused false negative PCR results for BTV3 at high RT temperature. Indeed, correction of both IVI-primers for BTV3 led to positive results for BTV3 but negative results for all other samples of the BT Proficiency Test 2020. Apparently, variability of the -3 position is sufficient for discriminative PCR detection, although the single mismatch in the IVI-reverse primer was the most important for this phenomenon. Extensive in silico validation showed that targets of both Seg-10 panBTV RT-PCR tests are not completely conserved, and the detailed effect of single mismatches are hard to predict. Therefore, we recommend at least two panBTV RT-PCR tests to minimize the risk of false negatives. Preferably, their PCR targets should be located at completely different and highly conserved regions of the BTV genome to guarantee adequate detection of future BTV infections.

Suspected SARS-Cov-2 reinfections in health care workers from Assam, India: Are they true reinfections?

Frontline healthcare workers (HCWs) are repeatedly exposed to SARS-CoV-2 and chance of exposure to it are invariably high than any other category of population. In this study, we investigated suspected cases of SARS-CoV-2 reinfection among eight HCWs involved in COVID-19 healthcare duty in Dibrugarh, Assam. Diagnosis of SARS-CoV-2 was done by Real Time RT-PCR or Rapid Antigen Detection Test at AMCH, Dibrugarh and ICMR-RMRC, Dibrugarh. Cases who tested positive for SARS-CoV-2 by RT-PCR or RAT for the second time and with symptoms suggestive of COVID-19 were included in this investigation as suspected cases of reinfection. SARS-CoV-2 IgG Ab titre and immune status ratio was estimated using a commercial SARS-CoV-2 Ig Ab detection ELISA kit. All eight cases were asymptomatic in first episode of infection with a C value above 30 and were non-reactive for SARS-CoV-2 IgG Ab. The second episode was symptomatic and marginally severe in some cases with C value less than 30 and with positive SARS-CoV-2 Ab titre. Most asymptomatic cases with C value above 30 failed to elicit immune response during the first episode. This may suggest that SARS-CoV-2 in low amount might be harbored transiently as bystander in droplet particles before being expelled from the nasal cavity which can be detected by the highly sensitive Real Time RT-PCR test. They may be below the infectious dose that is necessary to cause a clinical or a sub-clinical infection and fails to illicit an immune response. It is therefore very important to critically analyze the suspected cases of reinfection to be labelled as true reinfections. In conclusion, not all resurgence of symptoms with positive SARS-CoV-2 result for the second time after recovery are true reinfections and may be labelled as retest positives rather than reinfections. Further, routine surveillance of SARS-CoV-2 Ab testing for HCW is recommended to ascertain their immune status as they are the frontline workers of managing COVID-19 patients and are highly exposed to SARS-CoV-2 infection, and have a much higher risk of re-infection than the general public.

The period from prodromal fever onset to rash onset in laboratory-confirmed rubella cases: a cross-sectional study

BackgroundIn resource-limited settings, where rubella is endemic, it is difficult to determine which sporadic case should be tested for rubella. The study aimed to provide useful evidence to help screen rubella cases for real-time reverse transcriptase-polymerase chain reaction (RT-PCR) examination for rubella in resource-limited settings.MethodSuspected rubella patients identified by a physician and brought to the notice of the Ryugasaki public health center or the Tsuchiura public health center were enrolled from April 2018 through December 2019. The inclusion criterion was a confirmed rubella diagnosis based on laboratory tests. We studied the distribution of the time from the onset of fever until the onset of rash.ResultsThe study included 86 cases with simultaneous presentation of fever and rash. Twenty-nine cases had confirmed rubella based on the laboratory diagnosis. Among these, the time from the onset of fever until the onset of rash was limited to − 1 day to 2 days. The number of rubella cases was the highest when the onset of rash was on the following day of the onset of fever. Of the 78 patients who underwent the RT-PCR test, 48% tested positive for rubella among those with a time from the onset of fever to the onset of rash between − 1 day and 2 days (22 out of 46, 95% confidence interval 34–62%); no positive results (0 out of 30, 95% confidence interval - 14%) were seen in patients with a time from fever to rash onset ≥3 days.ConclusionThe period from the onset of fever to the onset of rash was limited to − 1 day to 2 days among confirmed rubella patients. If the period from onset of fever to the onset of rash was ≥3 days for a patient, the likelihood of rubella was low.

Comorbidities and risk factors for COVID-19 in a group of Iraqi patients confirmed by real-time PCR test

The aim of the study was to identify characteristic risk factors predisposing to coronavirus disease 2019 (COVID-19) outbreak in a group of Iraqi patients in Baghdad City. In this study, we included a total of 13411 persons who had been tested for COVID-19, supervised by Al-Razi Medical Center in Baghdad City during the period from June 20 to November 13, 2020. Cases were tested and divided into two groups according to the result of real-time RT-PCR test for COVID-19. A total of 1781 positive cases were detected with confirmed COVID-19. Comorbidities in the study patients were identified as hypertension, diabetes mellitus and respiratory disorders. Hypertension was the most common comorbidity in COVID-19 patients followed by diabetes mellitus. The results of this study showed that patients with comorbidities were significantly associated with positive RT-PCR results related to COVID-19. It was concluded that understanding the comorbidities enhance defining patients with COVID-19 at higher risk. Further research is needed to confirm and clarify the impact of the association of co-morbidities that accompany COVID-19.

Correlations between chest-CT and laboratory parameters in SARS-CoV-2 pneumonia: A single-center study from Italy.

To investigate the relationship between damaged lung assessed by chest computed tomography (CT) scan and laboratory biochemical parameters with the aim of finding other diagnostic tools.Patients who underwent chest CT for suspected Corona Virus Disease-2019 (COVID-19) pneumonia at the emergency department admission in the first phase of COVID-19 epidemic in Italy were retrospectively analyzed. Patients with both negative chest CT and absence of the novel coronavirus in nasopharyngeal or oropharyngeal real-time reverse transcriptase polymerase chain reaction (RT-PCR) swabs were excluded from the study. A total of 462 patients with positive CT scans for interstitial pneumonia were included in the study (250 males and 212 females, mean age 57 ± 17 years, range 18–89). Of these, 344 were positive to RT-PCR test, 118 were negative to double RT-PCR tests.CTs were analyzed for quantification of affected lung volume visually and by dedicated software. Statistical analysis to evaluate the relationship between laboratory analyses and CT patterns and amount of damaged lung related with COVID-19 pneumonia was performed in 2 groups of patients: positive RT-PCR COVID-19 group and negative RT-PCR COVID-19 group, but both with positive CT scans for interstitial pneumonia.Lymphocytopenia, C-reactive protein (CRP), lactate dehydrogenase (LDH), d-dimer, and fibrinogen increased levels occurred in most patients without statistically significant differences between the 2 groups with CT scans suggestive for COVID-19. In fact, in both groups the volume of lung damage was strongly associated with altered laboratory test results, even for patients with negative RT-PCR test.The decreased number of lymphocytes, and the increased levels of CRP, LDH, d-dimer, and fibrinogen levels are associated with SARS-CoV 2 related pneumonia. This may be useful as an additional diagnostic tool in patients with double negative RT-PCR assay and with highly suspected clinic and chest CT features for COVID-19 to isolate patients in a pandemic period.

Heart Transplantation Single Center Experience of 63 Heart Transplantation Procedures Performed during the COVID 19 Pandemic

PurposeThe COVID19 (C19) pandemic has resulted in a significant reduction in cardiac surgical procedures worldwide and in particular heart (OHT) transplantations. This was primarly due to increased risk of C19 infection and severity in immunocompromised patients (pts). Donor aviability decreased steeply, as intensive care units are struggling with C19 patients. Yet, number of patients requiring OHT increased rapidly, comprising of individuals with dreadful complications of untreated acute coronary syndromes.MethodsBetween March and October 2020 63 pts underwent OHT. The age range was 21-65 yrs and a median of 52 yr. All patients were evaluated by a multidisciplinary heart failure team (MDT), and placed on the HT waiting list once eligible. Once a donor has been identified, clinical history and examination obtained, a chest CT was taken and real time RTPCR test is ordered. Every recipient was subject to clinical screening, chest XRay, real RTPCR (24 hrs within HT)upon admission. All pts were looked after by experienced staff who tested negative for SARS-CoV-2, wearing PPE, working in isolation, single room care and visitors were not admitted.ResultsNo recipient or donor tested positive for SARS-CoV-2 at any stage in their care. 5 (8%) pts died, >30 days after the procedure, of multiorgan failure; all 5 pts were in INTERMACS II at the time of OHT. All patients were managed with our standard immunosuppression therapy consisting of mycofenolante mofetil and steroids in addition to calcineurin inhibitor (tacrolimus). According to recent results 25 patients (40%) had allograft rejection of ISHLT grade 3a detected by routine endomyocardial biopsies within first 3 months. The highdose steroids therapy was involved and positive response (ISHLT grade 0) was observed. 56 pts had a hospital stay of 30 d or less, 7 pts more than 30 d. Follow up visits were conducted using telemedicine unless it was absolutely necessary to see the pt in hospital. Also, we raised the threshold for endomyocardial biopsies by relying on clinical judgement and stable allograft function.ConclusionBy creating a robust system of care based on team work, early testing and retesting of patients and staff for SARSCoV2, the use of proper PPE's, adequate isolation and subsequent minimization of contact with pts, we were able to demonstrate that HT in the era of COVID19 pandemic can be performed safely.

Use of the COVID-19 Reporting and Data System (CO-RADS) classification and chest computed tomography involvement score (CT-IS) in COVID-19 pneumonia.

PurposeThe increasing tendency of chest CT usage throughout the COVID-19 epidemic requires new tools and a systematic scheme for diagnosing and assessing the lung involvement in Coronavirus Disease 2019 (COVID-19). To investigate the use of the COVID-19 Reporting and Data System (CO-RADS) classification and chest CT Involvement Score (CT-IS) in COVID-19 pneumonia.Material and methodsThis retrospective study enrolled 280 hospitalized patients diagnosed with COVID-19 pneumonia in a tertiary hospital in Turkey. All patients underwent non-contrast CT chest imaging. Two radiologists interpreted all CT images according to CO-RADS classification without knowing the clinical features, laboratory findings. We used CT involvement score (CT-IS) for assessing chest CT images of COVID-19 patients. Also, we examined the relationship between CT-IS and clinical outcomes in COVID-19 patients.ResultsOf the patients, 111(39.6%) had positive real-time reverse transcriptase-polymerase chain reaction (RT-PCR) results. CO-RADS 5 group patients had statistically significant positive RT-PCR results than the other groups (P < 0.001). All of the CO-RADS 2 group patients (30) had negative RT-PCR results. The mean total CT-IS in CO-RADS 2 group was 3.4 ± 2.8. The mean total CT-IS in CO-RADS 5 group was 8.2 ± 4.7. Total CT-IS was statistically significantly different among CO-RADS groups (P < 0.001). The mean total CT-IS was statistically significantly different between survivors and patients died of COVID-19 pneumonia (P < 0.001).ConclusionsCO-RADS is useful in detecting COVID-19 disease, even if RT-PCR testing is negative. CT-IS is also helpful as an imaging tool for evaluation of the severity and extent of COVID-19 pneumonia.

Clinical presentation and outcomes of the first patients with COVID-19 in Argentina: Results of 207079 cases from a national database.

There is limited evidence on the clinical characteristics of SARS-CoV-2 infection in Latin America. We present findings from a nationwide study in Argentina. What is disease severity measures and risk factors are associated with admission to an intensive care unit and mortality? Data were extracted from the COVID-19 database of the Integrated Argentina Health Information System, encompassing the period of March 3rd to October 2nd, 2020, using a standardized case report form that included information on contact history, clinical signs and symptoms, and clinical diagnosis. Information was collected at the initial site of care and follow-up conducted through calls by the regional healthcare authorities. A confirmed case of COVID-19 was defined as having a positive result through sequencing or real-time reverse-transcriptase polymerase chain reaction (RT-PCR) assay of nasal and pharyngeal swab specimens. RT-PCR testing was positive in 738,776 cases. Complete datasets were available for analysis in 207,079 cases. Mean age was 42.9±18.8 years, 50.0% were males. Frequent co-existing conditions included hypertension (19.2%), diabetes (9.7%), asthma (6.1%) and obesity (5.2%). Most common symptoms included fever (58.5%), cough (58.0%), headache (45.4%), and sore throat (42.1%). Death or ICU admission were independently associated with older age, male, coma, dyspnea or tachypnea, and seizures, with underlying co-morbidities such as immunodeficiency, chronic renal failure, and liver disease showing the strongest effects. Most cases of COVID-19 diagnosed in Argentina were mild and had a favorable outcome, but fatality rates were relatively elevated. Risk factors for adverse outcome included older age, male sex, coma and seizures, and the concurrent presence of several morbidities. These data may be useful for healthcare providers and healthcare policy makers of low-middle income and Latin American countries to guide decisions toward optimized care during the pandemic.

코로나바이러스감염증-19 진단검사에 대한 이해

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of coronavirus disease 2019 (COVID-19), is a type of human coronavirus that causes severe pneumonia, similar to SARS-CoV-1 and Middle East respiratory syndrome coronavirus. In Korea, the SARS-CoV-2 testing has started quickly from February 2020 to respond to the COVID-19 pandemic. In this article, I would like to introduce the characteristics of coronavirus and PCR test methods that play a large role in COVID-19 quarantine measures. Real-time reverse transcription (RT)-PCR is one of the molecular diagnostic method, and it detect SARS-CoV-2 RNA by amplifying SARS-CoV-2 specific RdRp (RNA-dependent RNA polymerase) gene and E (envelope) gene at the same time. Real-time RT-PCR is currently the most reliable test that confirming COVID-19 and is in use worldwide. Real-time RT-PCR test is recommended for COVID-19 confirmatory diagnosis in Korea, but this test requires dedicated equipment, reagents, experienced technicians and laboratory medicine specialists, and it takes about a few hours to a day to report. Rapid molecular testing results in one to two hours with a simple procedure. Antigen test is less sensitive than real-time RT-PCR and can only be used as a secondary role of diagnosis. As the global COVID-19 pandemic progresses, diagnostic testing guidelines and recommendations may vary and will be updated as scientific evidence and experience of the COVID-19 accumulates.

Feasibility and Performance of Self-Collected Nasal Swabs for Detection of Influenza Virus, Respiratory Syncytial Virus, and Human Metapneumovirus.

We assessed performance of participant-collected midturbinate nasal swabs compared to study staff-collected midturbinate nasal swabs for the detection of respiratory viruses among pregnant women in Bangkok, Thailand. We enrolled pregnant women aged ≥18 years and followed them throughout the 2018 influenza season. Women with acute respiratory illness self-collected midturbinate nasal swabs at home for influenza viruses, respiratory syncytial viruses (RSV), and human metapneumoviruses (hMPV) real-time RT-PCR testing and the study nurse collected a second midturbinate nasal swab during home visits. Paired specimens were processed and tested on the same day. The majority (109, 60%) of 182 participants were 20-30 years old. All 200 paired swabs had optimal specimen quality. The median time from symptom onsets to participant-collected swabs was 2 days and to staff-collected swabs was also 2 days. The median time interval between the 2 swabs was 2 hours. Compared to staff-collected swabs, the participant-collected swabs were 93% sensitive and 99% specific for influenza virus detection, 94% sensitive and 99% specific for RSV detection, and 100% sensitive and 100% specific for hMPV detection. Participant-collected midturbinate nasal swabs were a valid alternative approach for laboratory confirmation of influenza-, RSV-, and hMPV-associated illnesses among pregnant women in a community setting.

Inflammatory response in lungs and extrapulmonary sites detected by [18F] fluorodeoxyglucose PET/CT in convalescing COVID-19 patients tested negative for coronavirus

BackgroundThe severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has resulted in an ongoing global pandemic of coronavirus disease 2019 (COVID-19). The challenges associated with imaging infected patients have resulted, to date, in a paucity of metabolic imaging studies of patients with severe COVID-19 infection. Furthermore, it remains unclear if any abnormal metabolic events are taking place in patients who have recovered from COVID-19.PurposeTo use [18F] fluorodeoxyglucose ([18F] FDG) positron emission tomography/computed tomography (PET/CT) to measure metabolic activity in inflamed organs of patients convalescing post severe COVID-19 infection.Materials and methodsA prospective study was performed in seven convalescing patients who were recovering from severe COVID-19 infection in February 2020. Prior to [18F] FDG PET/CT, all patients had received two consecutive negative results of real-time reverse transcriptase polymerase chain reaction (RT-PCR) for SARS-CoV-2 nucleic acid. Clinical intake including symptoms, treatment, laboratory test results, and follow-up was performed. The PET/CT images of COVID-19 patients were compared to a control group of patients that were matched for age and sex.ResultsResidual pulmonary lesions were present in all patients and maximum standard uptake value (SUVmax), average standard uptake value (SUVavg), maximum CT intensity (CTmax), and average CT intensity (CTavg) were all significantly greater than in the control group (p < 0.01 for all). In addition, SUVmax and SUVavg were significantly greater in the mediastinal lymph node and liver, and SUVmax was significantly greater in the spleen, of COVID-19 patients compared with controls (p < 0.05 for all). For the spleen, SUVmax (r2 = 0.863, p = 0.003) and SUVavg (r2 = 0.797, p = 0.007) were significantly correlated with blood lymphocyte count, and which was below the normal range in five of the seven (71.4%) patients convalescing post severe COVID-19 infection.Conclusion[18F] FDG PET/CT quantitative analysis has shown that significant inflammation remained in lungs, mediastinal lymph nodes, spleen, and liver after two consecutive negative RT-PCR tests in patients convalescing post severe COVID-19 infection.

Pre-Operative SARS CoV-2 Rapid Antigen Test and Reverse Transcription Polymerase Chain Reaction: A conundrum in surgical decision making.

Purpose:Since the eye is in close proximity to the oro-nasal cavity, transmission of SARS CoV-2 is higher during ophthalmic surgical procedures, it is vital to ensure the safety of the healthcare team by pre-operative Rapid Antigen Test (RAT) and real-time Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) for SARS CoV-2 viral RNA of all patients planned for elective or emergency surgery. Hence, understanding the profile of pre-operative RAT and real-time RT-PCR among patients planned for various ophthalmic procedures, will help us make better decisions for future guidelines, for optimal planning of ophthalmic elective or emergency surgeries, keeping in mind the current scenario.Methods:This is a prospective descriptive study conducted on patients planned for elective or emergency ophthalmic surgeries, who had no COVID-19-related symptoms pre-operatively and were subjected to preoperative RAT and real-time RT-PCR of nasopharyngeal and throat swabs for SARS CoV-2 viral RNA. Data was compiled and statistically analysed.Results:204 patients underwent pre-operative RAT and RT-PCR testing; of which, 85 were females and 119 were males. Mean age of the study population was 51.44 ± 16.501 years. Among the 204, 2 tested positive for SARS CoV-2 with Rapid Antigen testing as well as RT-PCR and 10 patients tested positive via RT-PCR after a negative RAT result of the same sample.Conclusion:Pre-operative testing for COVID-19 disease is indispensable as there is a high chance of transmission from patient to healthcare workers. RAT has the advantage of quick results and may play a role in case of emergency procedures. However, it is prudent to perform the more sensitive real-time RT-PCR before any elective, non-emergency procedures or surgeries to avoid any undue risk to the healthcare team.

Diabetic Ketoacidosis and COVID-19: Two Case Reports

Background: COVID-19 is a viral infection that causes pneumonia with dyspnea, cough, and fever. Its outcomes are more severe in patients with diabetes, hypertension, and other disorders than in healthy people. New-onset diabetes and diabetic ketoacidosis (DKA) have been reported as the complications of COVID-19 in several studies. It seems that the prevalence of DKA due to Covid-19 is increasing. Therefore, two cases of individuals with DKA due to COVID-19 are presented in this study to inform other researchers about the details of this phenomenon. Material and Methods: Demographic characteristics, medical histories, physical examinations, laboratory investigations, real-time RT-PCR tests, computed tomography (CT) imaging studies, given treatments, clinical courses, and management outcomes were documented prospectively. Results: In the present experience, the manifestation of COVID-19 disease in the second case with a history of diabetes was more severe than in the first case. In the second case, with underlying diabetes and COVID-19, the DKA manifestation was associated with consciousness loss, severe restlessness, and respiratory distress;however, in the first case, with COVID-19 without diabetes, the DKA manifestation was associated with anorexia, weight loss, and lack of respiratory distress, which were milder symptoms than the second case. Conclusion: Due to the high prevalence of diabetes mellitus and COVID-19 in Iran, it is recommended to raise awareness of DKA symptoms among health professionals. © The Author(s) 2021;All rights reserved. Published by Rafsanjan University of Medical Sciences Press.

Relationship of Hypocalciamia and Severity of Disease in COVID-19 Patients in Lahore Pakistan

Introduction: In Pakistan, a wave of coronavirus has started in February which was first reported in Wahun City China. Abnormal calcium blood level is considered one of the important and common electrolyte disturbances in patients with severe infection. The effect of blood calcium level on the severity of COVID-19 and the possible immune regulation mechanism is considered to be an important factor related to severity of disease. Aim of the Study: To find out the correlation of blood calcium levels and related risk factors of patients suffering from COVID-19 and provide a basis for early intervention in patients with relevant clinical characteristics and further reduce the incidence of critical illness and mortality in COVID-19 patients. Method: 180 COVID-19 patients were admitted to Gulab Devi Teaching Hospital, Lahore between February to July 2020. Subject index were tested on real-time fluorescent RT-PCR test. On the basis of the result of RT-PCR, they were divided into two groups according to clinical symptoms as mild (control/placebo group) and critical (experimental group). The patient with the mild symptom were included in the placebo group, whereas patient with the severe symptoms were included in the experimental group. Exclusion criteria: The patients who do not show the detection level of viral infection with the RT-PCR are excluded from the study. Results: The comparison has been carried out between the placebo group and the experimental group on the basis of various parameters. The patients serum calcium level was tested and the value for hypocalcemia was declared as per universal recommendation which is < 2.13mmol/L. According to the corrected serum total calcium level, COVID-19 patients can be divided into hypocalcemia group and normal blood calcium group. The result showed that hypocalcemia and immune dysfunction was found high in experimental group as compare to control group which shows a strong correlation between hypocalcemia and COVID-19 disease. Conclusion: It has been concluded from the study that patients with COVID-19 have obvious hypocalcemia and immune dysfunction, especially in experimental group patients. In this study, it is easy to be combined with multiple infections caused by pathogenic bacteria under certain conditions. Close monitoring of blood calcium levels can predict the severity of the disease more effectively.