scholarly journals Study of vertical transmission of COVID-19 infection in COVID-19 positive obstetrical patients by comparing amniotic fluid and immediate neonatal COVID-19 RT-PCR

2021 ◽  
Vol 10 (9) ◽  
pp. 3383
Author(s):  
Smita Tyagi ◽  
Rajesh Kumar Agarwal ◽  
Manish Agarwal ◽  
Manika Kapoor ◽  
Sakshi Tiwari
Keyword(s):  
Amniotic Fluid ◽  
Vertical Transmission ◽  
Neonatal Death ◽  
Rt Pcr ◽  
Normal Delivery ◽  
Medical College ◽  
Viral Transport ◽  
Viral Transport Medium ◽  
Oropharyngeal Swab ◽  
Medium Sample

Background: The objective of this study was to estimate the incidence of vertical transmission COVID-19 by RT-PCR.Methods: In this hospital based prospective study, all COVID-19 positive pregnant women admitted in COVID ward in Muzaffarnagar medical college and hospital from April 2020 to January 2021 were included. A detailed history and examination was done and all routine investigations were done as per protocol. Samples were taken from amniotic fluid during vaginal delivery or caesarean section and collected in viral transport medium. Sample were also collected as nasal and oropharyngeal swab from neonate immediately after birth and sent for COVID 19 RT-PCR.Results: Out of total 50 cases; 43 (86%) neonates were delivered via LSCS and 7 (14%) by normal delivery, out of these 41 (82%) neonates were normal; 4 (8%) were IUGR, 2 (4%) were IUD, 2 (4%) were pre-term and 1 (2%) neonatal death. All 50 amniotic fluid as well as nasal and oropharyngeal samples of all neonates were negative by RT-PCR.Conclusions: Low vertical transmission may be due to the fact that antibodies are produced by mother that crosses the placenta and saves the fetus or there are highly specific immunological mediators in the placenta that do not allow the infection to pass to the fetus.

scholarly journals Sensitivity of nasopharyngeal, oropharyngeal and nasal washes specimens for SARS-CoV-2 detection in the setting of sampling device shortage

2020 ◽  
Author(s):  
Adrien Calame ◽  
Lena Mazza ◽  
Adriana Renzoni ◽  
Laurent Kaiser ◽  
Manuel Schibler
Keyword(s):  
Culture Medium ◽  
Alternative Methods ◽  
Analytical Sensitivity ◽  
Sample Collection ◽  
Rt Pcr ◽  
Sampling Device ◽  
Viral Transport ◽  
Viral Transport Medium ◽  
A Cell ◽  
Oropharyngeal Swab

In the context of an unprecedented shortage of nasopharyngeal swabs (NPS) or sample transport media during the coronavirus disease 2019 (COVID-19) crisis, alternative methods for sample collection are needed. To address this need, we validated a cell culture medium as a viral transport medium, and compared the analytical sensitivity of SARS-CoV-2 real-time RT-PCR in nasal wash (NW), oropharyngeal swab (OPS) and NPS specimens. Both the clinical and analytical sensitivity were comparable in these three sample types. OPS and NW specimens may therefore represent suitable alternatives to NPS for SARS-CoV-2 detection.

scholarly journals Extraction Free Rapid Detection of SARS-CoV-2 from Oropharyngeal/Nasopharyngeal Swabs by Real Time PCR

2021 ◽  
Author(s):  
Parul Sinha ◽  
Sandeep Gupta ◽  
Megha Gupta ◽  
Dinesh Kumar Jain ◽  
Malvika Sharma ◽  
...  
Keyword(s):  
Real Time ◽  
Extraction Method ◽  
Rna Extraction ◽  
Molecular Testing ◽  
Acid Extraction ◽  
Rt Pcr ◽  
Transport Medium ◽  
Viral Transport ◽  
Viral Transport Medium ◽  
Medium Sample

Introduction: The emergence of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) pandemic has been troublesome particularly for developing countries that lack infrastructure and capacities to produce the kits locally. Simplification of the method can increase diagnostic efficiency which can benefit patients and help in infection control, consequently saving time and lives. Aim: To evaluate the diagnostic value of four methods (that omit extraction step) for detection of SARS-CoV-2 against the traditional extraction method. Materials and Methods: This was a cross-sectional analysis for evaluating diagnostic accuracy of four methods for detection of SARS-CoV-2 by real-time Reverse Transcriptase Polymerase Chain Reaction (rRT-PCR), conducted in the Department of Microbiology, SMS Medical College, Jaipur, Rajasthan, India, in October 2020. Ninety four SARS-CoV-2 RT-PCR positive samples and 20 negative samples were taken for this study. Automated extraction system was used for Ribonucleic Acid (RNA) extraction and four different approaches were compared to the traditional extraction method for detection of SARS-CoV-2 by RT-PCR. Data was entered and analysed using Statistical Package for the Social Sciences (SPSS) statistical software version 24.0. Results: The automated RNA extraction method was compared to the method of direct addition of samples with (Heat processed Direct Viral transport medium Sample (HDVS)) and without heating (Direct Viral transport medium Sample (DVS)), directs addition of diluted (1:5) sample with (Heat processed diluted VTM sample (HdVS)) and without heating (Diluted VTM sample (dVS)) as well as after addition of Proteinse K (PK) to the diluted samples that came either negative/invalid. Out of four methods, the HdVS method gave the best results, considering extraction with Perkin Elmer as standard, this method showed sensitivity of 96.74%, specificity of 100%. Conclusion: In current pandemic, molecular testing is critically challenged by the limited supplies of reagents of nucleic acid extraction alternative method like diluting and heating of Viral Transport Media (VTM) samples and using them directly as elutes serve as an easy, fast and inexpensive alternative.

scholarly journals Vertical Transmission of Severe Acute Respiratory Syndrome Coronavirus 2: A Systematic Review

2020 ◽  
Vol 37 (10) ◽  
pp. 1055-1060 ◽  
Author(s):  
Ziyi Yang ◽  
Yi Liu
Keyword(s):  
Systematic Review ◽  
Severe Acute Respiratory Syndrome ◽  
Amniotic Fluid ◽  
Cord Blood ◽  
Vertical Transmission ◽  
Meta Analysis ◽  
Nasopharyngeal Swab ◽  
Rt Pcr ◽  
Serum Samples ◽  
Virus Specific Antibody

Objective The aim of this study is to summarize currently available evidence on vertical transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Study Design A systematic review was conducted following the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-analysis Statement. Results A total of 22 studies comprising 83 neonates born to mothers diagnosed with coronavirus disease 2019 were included in the present systematic review. Among these neonates, three were confirmed with SARS-CoV-2 infection at 16, 36, and 72 hours after birth, respectively, by nasopharyngeal swab real-time polymerase chain reaction (RT-PCR) tests; another six had elevated virus-specific antibody levels in serum samples collected after birth, but negative RT-PCR test results. However, without positive RT-PCR tests of amniotic fluid, placenta, or cord blood, there is a lack of virologic evidence for intrauterine vertical transmission. Conclusion There is currently no direct evidence to support intrauterine vertical transmission of SARS-CoV-2. Additional RT-PCR tests on amniotic fluid, placenta, and cord blood are needed to ascertain the possibility of intrauterine vertical transmission. For pregnant women infected during their first and second trimesters, further studies focusing on long-term outcomes are needed. Key Points

scholarly journals Vertical transmission of Severe Acute Respiratory Syndrome Coronavirus 2: A scoping review

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250196
Author(s):  
Lemi Belay Tolu ◽  
Alex Ezeh ◽  
Garumma Tolu Feyissa
Keyword(s):  
Breast Milk ◽  
Severe Acute Respiratory Syndrome ◽  
Amniotic Fluid ◽  
Cord Blood ◽  
Pregnant Women ◽  
Vertical Transmission ◽  
Throat Swab ◽  
Rt Pcr ◽  
Third Trimester ◽  
Adequate Sample Size

Introduction The evidence for vertical transmission of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is not well established. Therefore, the objective of this review is to summarize emerging evidence on the vertical transmission of Severe Acute Respiratory Syndrome Coronavirus 2. Methods We conducted a systematic search in PubMed, CINAHL, Web of Science, SCOPUS, and CENTRAL. Likewise, a search for preprint publications was conducted using MedRxiv and Research Square. Studies that addressed vertical transmission of SARS-CoV-2 (concept) among pregnant women infected by Covid-19 (population) in any setting (community, hospital, or home) in any country or context were considered for inclusion. Any types of studies or reports published between December 2019 and September 2020 addressing the effects of SARS-CoV-2 on pregnant women and their newborn babies were included. Studies were screened for eligibility against the inclusion criteria for the review by two reviewers. Results We identified 51 studies reporting 336 newborns screened for COVID-19. From the 336 newborns screened for COVID-19, only 15 (4.4%) were positive for throat swab RT-PCR. All neonates with positive throat swab RT-PCR were delivered by cesarean section. Among neonates with throat swab SARS-CoV-2 positive only five (33.3%) had concomitant placenta, amniotic fluid, and cord blood samples tested, of which only one amniotic fluid sample is positive for RT PCR. Five neonates had elevated IgG and IgM but without intrauterine tissue tested. Four neonates had chest imaging suggestive of COVID-19 pneumonia. Conclusion Currently there is not enough evidence on vertical virologic transmission of COVID-19 infection during the third trimester of pregnancy. Additionally, there is no evidence to support cesarean delivery, abstaining from breast feeding nor mother and infant separation. Further research involving an adequate sample size of breast milk, placenta, amniotic fluid, and cord blood to ascertain the possibility of vertical transmission and breast milk transfer is needed.

scholarly journals Sensitivity of nasopharyngeal, oropharyngeal, and nasal wash specimens for SARS-CoV-2 detection in the setting of sampling device shortage

2020 ◽  
Author(s):  
Adrien Calame ◽  
Léna Mazza ◽  
Adriana Renzoni ◽  
Laurent Kaiser ◽  
Manuel Schibler
Keyword(s):  
Culture Medium ◽  
Alternative Methods ◽  
Analytical Sensitivity ◽  
Sample Collection ◽  
Sampling Device ◽  
Nasal Wash ◽  
Viral Transport ◽  
Viral Transport Medium ◽  
A Cell ◽  
Oropharyngeal Swab

Abstract In the context of an unprecedented shortage of nasopharyngeal swabs (NPS) or sample transport media during the coronavirus disease 2019 (COVID-19) crisis, alternative methods for sample collection are needed. To address this need, we validated a cell culture medium as a viral transport medium, and compared the analytical sensitivity of SARS-CoV-2 RT-PCR in nasal wash (NW), oropharyngeal swab (OPS), and NPS specimens. Both the clinical and analytical sensitivity were comparable in these three sample types. OPS and NW specimens may therefore represent suitable alternatives to NPS for SARS-CoV-2 detection.

scholarly journals In-House Viral Transport Medium (VTM) Manufacture in the Time of Shortage, Supply and Crisis of COVID-19 at Veteran Affairs Medical Center (VAMC)

2020 ◽  
Vol 154 (Supplement_1) ◽  
pp. S161-S162
Author(s):  
J M Petersen ◽  
S Dalal ◽  
D Jhala
Keyword(s):  
Amphotericin B ◽  
Medical Center ◽  
Phenol Red ◽  
Rt Pcr ◽  
Gentamicin Sulfate ◽  
Viral Transport ◽  
Viral Transport Medium ◽  
Fetal Bovine ◽  
National Emergency ◽  
Negative Controls

Abstract Introduction/Objective With the outbreak of COVID-19 caused by SARS-CoV-2, there have been challenges in the maintenance of adequate supplies both in terms of PPE and for testing. The shortage of commercial VTM for the transport of specimens for PCR testing has created a situation in which laboratories would need to manufacture their own in-house VTM as commonly used commercial VTM is unavailable. However, there is sparse literature on the emergency manufacture of VTM. Here, we describe the VAMC experience in manufacturing/quality control on its own VTM. Results VTM was manufactured by pathology and laboratory medicine using strict aseptic technique with Hanks Balanced Salt Solution (HBSS) 500 ml bottle with phenol red, sterile heat-inactivated fetal bovine serum (FBS) gentamicin sulfate (50 mg/mL) and amphotericin B (250 ug/ml). First, 50 ml of amphotericin B and 50 ml of gentamicin sulfate were mixed. Then 10 ml of FBS was mixed with the HBSS bottle and then 2 ml of the gentamicin/amphotericin B mixture was also mixed into the HBSS bottle. 3 ml aliquots were made from this mixture to constitute individual tubes of VTM for clinical use. Sterility for each batch (after 24-hour incubation at 37o C in the CO2 incubator) was assessed visually and by culture on a blood agar, chocolate agar, and thioglycolate mediums. An efficacy check was performed for each batch by spiking positive and negative controls into the VTM aliquots; RT-PCR for SARS-CoV-2 was executed to verify the medium did not degrade viral RNA and produced expected results for room temperature, refrigerated, and frozen samples. Previously manufactured VTM without phenol red also underwent sterility and efficacy checks. Results VTM was successfully manufactured in-house, allowing testing to continue despite the shortage. Sterility and efficacy checks on all lots and bottles from which the VTM aliquots were made passed with no growth detected and efficacy passing with all expected positives and negatives resulting as expected. Conclusion To the author’s knowledge, this represents the first published abstract on VTM manufacture in this most unprecedented crisis involving COVID-19. In this national emergency with corresponding shortage of testing supplies including commercial VTM, the in-house manufacture of VTM is both feasible and prudent to ensure continuity of testing and quality patient/laboratory care.

scholarly journals Development and Clinical Evaluation of an Immunochromatography-Based Rapid Antigen Test (GenBody™ COVAG025) for COVID-19 Diagnosis

Viruses ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 796
Author(s):  
Doyeong Kim ◽  
Jihoo Lee ◽  
Jyotiranjan Bal ◽  
Seul Ki Seo ◽  
Chom-Kyu Chong ◽  
...  
Keyword(s):  
Real Time ◽  
Sensitivity And Specificity ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Clinical Sensitivity ◽  
Viral Transport ◽  
Global Pandemic ◽  
Viral Transport Medium ◽  
Antigen Tests ◽  
Higher Sensitivity

Antigen tests for SARS-CoV-2 diagnosis are simpler and faster than their molecular counterparts. Clinical validation of such tests is a prerequisite before their field applications. We developed and clinically evaluated an immunochromatographic immunoassay, GenBody™ COVAG025, for the rapid detection of SARS-CoV-2 nucleocapsid (NP) antigen in two different clinical studies. Retrospectively, 130 residual nasopharyngeal swabs transferred in viral transport medium (VTM), pre-examined for COVID-19 through emergency use authorization (EUA)-approved real-time RT-PCR assay and tested with GenBody™ COVAG025, revealed a sensitivity and specificity of 90.00% (27/30; 95% CI: 73.47% to 97.89%) and 98.00% (98/100; 95% CI: 92.96% to 99.76%), respectively, fulfilling WHO guidelines. Subsequently, the prospective examination of 200 symptomatic and asymptomatic nasopharyngeal swabs, collected on site and tested with GenBody™ COVAG025 and EUA-approved real-time RT-PCR assay simultaneously, revealed a significantly higher sensitivity and specificity of 94.00% (94/100; 95% CI: 87.40% to 97.77%) and 100.00% (100/100; 95% CI: 96.38% to 100.00%), respectively. Clinical sensitivity and specificity were significantly high for samples with Ct values ≤ 30 as well as within 3 days of symptom onset, justifying its dependency on the viral load. Thus, it is assumed this can help with the accurate diagnosis and timely isolation and treatment of patients with COVID-19, contributing to better control of the global pandemic.

scholarly journals Evaluation of Saline as Potential Alternative to Viral Transport Media for COVID-19 Samples Stored at Different Temperatures

2021 ◽  
Author(s):  
Parul Sinha ◽  
Dinesh Kumar Jain ◽  
Sandeep Gupta ◽  
Monika Gupta ◽  
Megha Gupta ◽  
...  
Keyword(s):  
Normal Saline ◽  
Effect Of Temperature ◽  
Nucleic Acid Detection ◽  
Infected People ◽  
Viral Transport ◽  
Viral Transport Medium ◽  
Different Temperatures ◽  
Oropharyngeal Swab ◽  
Potential Alternative ◽  
The Cost

Introduction: Severe acute respiratory syndrome coronavirus-2 (SARS CoV 2) virus, a causative agent of COVID-19 has led to universal pandemic. During this pandemic there has been an acute shortage of good quality Viral Transport Medium (VTM) because of increase in number of infected people worldwide. It is also difficult to maintain the transport and storing conditions in line with the guidelines in pandemics. Aim: To assess the feasibility of Oropharyngeal Swab (OP)/Nasal swabs in 0.9% normal saline in place of VTM and to analyse the effect of temperature on nucleic acid detection by rRT PCR on saline samples stored at 4ºC, ambient and at higher temperature (37ºC). Materials and Methods: The present study was an observational analytical study which included 94 positive and 5 negative samples. Patients' nasal or OP samples were collected as dry swabs and in VTM. Normal saline was added once the samples were received in the laboratory. PCR was done with saline and VTM samples both on day 1. Samples were aliquotted in 3 sets and one set was kept at 4º-8º C and other two at 25ºC and 37ºC, respectively. All positive samples were further tested on day 3, day 4 and day 6. Results were analysed and compared. Results: Samples in normal saline showed very good sensitivity at all temperatures (4º-8ºC, 25ºC and 37ºC) till day 6. Both the swab samples (in saline and in VTM) showed nearly 100% agreement in rRT-PCR results. Ct value variation was also ≤±2. Conclusion: Looking into the cost and logistics issues especially during pandemics, saline is a good and cheaper alternative to VTM and with its use, testing capacity can be expanded.

Effective optimization of SARS-CoV-2 laboratory testing variables in an era of supply chain constraints

2020 ◽  
Vol 15 (15) ◽  
pp. 1483-1487
Author(s):  
Nikhil S Sahajpal ◽  
Ashis K Mondal ◽  
Allan Njau ◽  
Sudha Ananth ◽  
Kimya Jones ◽  
...  
Keyword(s):  
Supply Chain ◽  
Rna Extraction ◽  
Nasopharyngeal Swab ◽  
Sample Collection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Short Supply ◽  
Viral Transport ◽  
3D Printed ◽  
Bridging Studies

RT-PCR-based assays for the detection of SARS-CoV-2 have played an essential role in the current COVID-19 pandemic. However, the sample collection and test reagents are in short supply, primarily due to supply chain issues. Thus, to eliminate testing constraints, we have optimized three key process variables: RNA extraction and RT-PCR reactions, different sample types and media to facilitate SARS-CoV-2 testing. By performing various validation and bridging studies, we have shown that various sample types such as nasopharyngeal swab, bronchioalveolar lavage and saliva, collected using conventional nasopharyngeal swabs, ESwab or 3D-printed swabs and, preserved in viral transport media, universal transport media, 0.9% sodium chloride or Amies media are compatible with RT-PCR assay for COVID-19. Besides, the reduction of PCR reagents by up to fourfold also produces reliable results.

scholarly journals Diagnosis of SARS-Cov-2 Infection by RT-PCR Using Specimens Other Than Naso- and Oropharyngeal Swabs: A Systematic Review and Meta-Analysis

Diagnostics ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 363
Author(s):  
Vânia M. Moreira ◽  
Paulo Mascarenhas ◽  
Vanessa Machado ◽  
João Botelho ◽  
José João Mendes ◽  
...  
Keyword(s):  
Systematic Review ◽  
Clinical Performance ◽  
Point Of Care ◽  
Meta Analysis ◽  
High Sensitivity ◽  
Nasopharyngeal Swab ◽  
Sample Collection ◽  
Rt Pcr ◽  
Oropharyngeal Swab ◽  
Nucleic Acid Assays

The rapid and accurate testing of SARS-CoV-2 infection is still crucial to mitigate, and eventually halt, the spread of this disease. Currently, nasopharyngeal swab (NPS) and oropharyngeal swab (OPS) are the recommended standard sampling techniques, yet, these have some limitations such as the complexity of collection. Hence, several other types of specimens that are easier to obtain are being tested as alternatives to nasal/throat swabs in nucleic acid assays for SARS-CoV-2 detection. This study aims to critically appraise and compare the clinical performance of RT-PCR tests using oral saliva, deep-throat saliva/posterior oropharyngeal saliva (DTS/POS), sputum, urine, feces, and tears/conjunctival swab (CS) against standard specimens (NPS, OPS, or a combination of both). In this systematic review and meta-analysis, five databases (PubMed, Scopus, Web of Science, ClinicalTrial.gov and NIPH Clinical Trial) were searched up to the 30th of December, 2020. Case-control and cohort studies on the detection of SARS-CoV-2 were included. The methodological quality was assessed using the Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS 2). We identified 1560 entries, 33 of which (1.1%) met all required criteria and were included for the quantitative data analysis. Saliva presented the higher accuracy, 92.1% (95% CI: 70.0–98.3), with an estimated sensitivity of 83.9% (95% CI: 77.4–88.8) and specificity of 96.4% (95% CI: 89.5–98.8). DTS/POS samples had an overall accuracy of 79.7% (95% CI: 43.3–95.3), with an estimated sensitivity of 90.1% (95% CI: 83.3–96.9) and specificity of 63.1% (95% CI: 36.8–89.3). The remaining index specimens could not be adequately assessed given the lack of studies available. Our meta-analysis shows that saliva samples from the oral region provide a high sensitivity and specificity; therefore, these appear to be the best candidates for alternative specimens to NPS/OPS in SARS-CoV-2 detection, with suitable protocols for swab-free sample collection to be determined and validated in the future. The distinction between oral and extra-oral salivary samples will be crucial, since DTS/POS samples may induce a higher rate of false positives. Urine, feces, tears/CS and sputum seem unreliable for diagnosis. Saliva testing may increase testing capacity, ultimately promoting the implementation of truly deployable COVID-19 tests, which could either work at the point-of-care (e.g. hospitals, clinics) or at outbreak control spots (e.g., schools, airports, and nursing homes).

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