scholarly journals Optimal Preparation of COVID-19 Viral Transport Medium for Culture

2020 ◽  
Author(s):  
Julie McAuley ◽  
Claire Fraser ◽  
Elena Paraskeva ◽  
Elizabeth Trajcevska ◽  
Michelle Sait ◽  
...  
Keyword(s):  
Diagnostic Testing ◽  
Vero Cells ◽  
Nucleic Acid Detection ◽  
Transport Medium ◽  
Viral Culture ◽  
Viral Transport ◽  
Viral Transport Medium ◽  
Escape Mutants ◽  
Diagnostic Approaches ◽  
Manual Handling

Abstract IntroductionThe sudden arrival of the COVID-19 pandemic placed significant stresses on supply chains including viral transport medium (VTM). The VTM that was urgently required needed to support viral replication, as well as other routine diagnostic approaches. We describe the preparation and validation testing of VTM for rapidly expanding diagnostic testing, where the capacity of the VTM to preserve viral integrity, for culture, isolation and full sequence analysis, was maintained.MethodsVTM was prepared using different methods of sterilization then ‘spiked’ with virus. The VTM was investigated using viral culture in Vero cells, and for nucleic acid detection by quantitative PCR (qPCR).ResultsThe best results were obtained by filter and autoclave-based sterilization. The VTM proved robust for culture-based analyses provided the inoculated VTM was stored at 4oC, and tested within 48 hours. The filtered VTM also supported PCR-based diagnosis for at least 5 days when the mock inoculated VTM was held at room temperature (RT).DiscussionThe manual handling of VTM production, including filling and sterilization, was optimized. SARS-CoV-2 was spiked into VTM to assess different sterilization methods and measure the effects of storage time and temperature upon VTM performance. While most diagnostic protocols will not require replication competent virus, the use of high quality VTM will allow for the next phase of laboratory analysis in the COVID-19 pandemic, including drug and antibody susceptibility analysis of re-isolated SARS--CoV-2, and for the testing of vaccine escape mutants.

scholarly journals Optimal preparation of SARS-CoV-2 viral transport medium for culture

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Julie McAuley ◽  
Claire Fraser ◽  
Elena Paraskeva ◽  
Elizabeth Trajcevska ◽  
Michelle Sait ◽  
...  
Keyword(s):  
Diagnostic Testing ◽  
Vero Cells ◽  
Nucleic Acid Detection ◽  
Transport Medium ◽  
Viral Culture ◽  
Viral Transport ◽  
Viral Transport Medium ◽  
Escape Mutants ◽  
Diagnostic Approaches ◽  
Manual Handling

Abstract Introduction The sudden arrival of the COVID-19 pandemic placed significant stresses on supply chains including viral transport medium (VTM). The VTM that was urgently required needed to support viral replication, as well as other routine diagnostic approaches. We describe the preparation and validation testing of VTM for rapidly expanding diagnostic testing, where the capacity of the VTM to preserve viral integrity, for culture, isolation and full sequence analysis, was maintained. Methods VTM was prepared using different methods of sterilization then ‘spiked’ with virus. The VTM was investigated using viral culture in Vero cells, and for nucleic acid detection by quantitative PCR. Results The best results were obtained by filter and autoclave-based sterilization. The VTM proved robust for culture-based analyses provided the inoculated VTM was stored at 4 °C, and tested within 48 h. The filtered VTM also supported PCR-based diagnosis for at least 5 days when the mock inoculated VTM was held at room temperature. Discussion The manual handling of VTM production, including filling and sterilization, was optimized. SARS-CoV-2 was spiked into VTM to assess different sterilization methods and measure the effects of storage time and temperature upon VTM performance. While most diagnostic protocols will not require replication competent virus, the use of high quality VTM will allow for the next phase of laboratory analysis in the COVID-19 pandemic, including drug and antibody susceptibility analysis of re-isolated SARS-CoV-2, and for the testing of vaccine escape mutants.

scholarly journals Dry Swab Method of sample collection for SARS-CoV2 testing can be used for culturing virus

2021 ◽  
Author(s):  
Sushma Ram ◽  
M. Ghalib Enayathullah ◽  
Yash Parekh ◽  
Karthik Bharadwaj Tallapaka ◽  
Rakesh K Mishra ◽  
...  
Keyword(s):  
Room Temperature ◽  
Rna Extraction ◽  
Vero Cells ◽  
Sample Collection ◽  
Qpcr Data ◽  
Transport Medium ◽  
Viral Transport ◽  
Viral Particles ◽  
Extraction Step ◽  
Viral Transport Medium

Background: Earlier studies suggested the use of dry swab method for SARS-CoV-2 detection as it does not need VTM and subsequent RNA extraction step making the process cheaper, safer and faster. In this study we explore whether the virus in the dry swab is viable and can be cultured and propagated. Method: Swabs were spiked with SARS-CoV-2 and stored in three different conditions: a) as dry swab (SD, eluted in 1 mL DMEM), b) in 1 mL of Viral Transport Medium (SVTM), and c) in 1 mL of Tris-EDTA buffer (STE). The sample groups were stored either at room temperature (RT ,25°C±1°C) or at 4°C for 1, 4, 8, 12, 24, 48 and 72 hours before being used as viral inoculums for the propagation studies in Vero cells. Results: The RT-qPCR data suggests that SD incubated both at RT and 4°C harbors viral particles that are viable and culturable at par with SVTM and STE. Conclusion: The dry swab method, in addition to its advantages in detection of the virus, also renders viable viral particles that can be cultured and propagated.

scholarly journals Studi Awal Analisis Molekuler Human Papillomavirus dari Apusan Glans dan Batang Penis

2021 ◽  
Vol 9 (4) ◽  
pp. 433
Author(s):  
Maya Savira ◽  
Resty Yuwandari ◽  
Yossi Maryanti ◽  
Rahmat Azhari Kemal ◽  
Donel S
Keyword(s):  
Human Papillomavirus ◽  
Rna Extraction ◽  
Viral Rna ◽  
Transport Medium ◽  
Viral Transport ◽  
Viral Transport Medium

Pria juga dapat mengalami keganasan akibat infeksi Human Papillomavirus (HPV) serta bertindak sebagai reservoir virus. Metode skrining HPV pada wanita telah terstandardisasi, namun belum ada standar metode skrining pada pria di Indonesia. Beberapa studi pada populasi pria di luar negeri menunjukkan potensi sampling pada daerah genitalia eksterna untuk skrining HPV. Tujuan: mengoptimasi metode skrining HPV secara molekuler pada pria. Metode: Responden adalah partner seksual wanita pansien kanker serviks di RSUD Arifin Achmad Provinsi Riau. Apusan dari glans dan batang penis diambil menggunakan nylon-flocked swab yang kemudian dimasukkan ke dalam 350µl viral transport medium terpisah. DNA diisolasi dari sampel yang kemudian dianalisis untuk mendeteksi gen human β-globin dan HPV. Hasil: Optimasi awal menunjukkan gen β-globin dapat terdeteksi dari hasil ekstraksi dengan kit Zeesan Viral RNA Extraction. Pita HPV hasil PCR dengan primer MY09 dan MY11 dapat muncul namun masih tipis. Simpulan: Studi awal ini menunjukkan bahwa apusan glans dan batang penis dapat digunakan untuk deteksi HPV secara molekuler pada pria, namun proses pengambilan sampel, ekstraksi DNA, dan PCR masih perlu dioptimasi.Kata kunci: apusan, glans, HPV, penis

scholarly journals SARS-CoV-2 detection in setting of viral swab scarcity: are MRSA swabs and viral swabs equivalent?

2020 ◽  
Author(s):  
Daniel G. Federman ◽  
Shaili Gupta ◽  
Gary Stack ◽  
Sheldon M. Campbell ◽  
David R. Peaper ◽  
...  
Keyword(s):  
Nasal Swab ◽  
High Rate ◽  
Test Results ◽  
Sample Collection ◽  
Transport Medium ◽  
Viral Transport ◽  
Global Pandemic ◽  
Viral Transport Medium ◽  
Nasal Swabs ◽  
Viral Media

AbstractBackgroundThe global pandemic of Severe Acute Respiratory Syndrome-Related Coronavirus 2 (SARS-CoV2) has resulted in unprecedented challenges for healthcare systems. One barrier to widespread testing has been a paucity of traditional respiratory viral swab collection kits relative to the demand. Whether other sample collection kits, such as widely available MRSA nasal swabs can be used to detect SARS-CoV-2 is unknown.MethodsWe compared simultaneous nasal MRSA swabs (COPAN ESwabs ® 480C flocked nasal swab in 1mL of liquid Amies medium) and virals wabs (BD H192(07) flexible mini-tip flocked nasopharyngeal swabs in 3mL Universal Transport Medium) for SARS-CoV-2 PCR testing using Simplexa COVID-19 Direct assay on patients over a 4-day period. When the results were discordant, the viral swab sample was run again on the Cepheid Xpert Xpress ® SARS-CoV-2 assay.ResultsOf the 81 included samples, there were 19 positives and 62 negatives in viral media and 18 positives and 63 negative in the MRSA swabs. Amongst all included samples, there was concordance between the COPAN ESwabs ® 480C and the viral swabs in 78 (96.3%).ConclusionWe found a high rate of concordance in test results between COPAN ESwabs ® 480C in Amies solution and BD H192(07) nasopharyngeal swabs in in 3 mL of Universal Viral Transport medium viral media. Clinicians and laboratories should feel better informed and assured using COPAN ESwabs ® 480C to help in the diagnosis of COVID-19.

Stability of different viruses in a newly developed transport medium

1974 ◽  
Vol 20 (1) ◽  
pp. 75-80 ◽  
Author(s):  
F. R. Bishai ◽  
N. A. Labzoffsky
Keyword(s):  
Cerebrospinal Fluid ◽  
Herpes Simplex ◽  
Serum Proteins ◽  
The Other ◽  
Autopsy Material ◽  
Herpes Simplex Viruses ◽  
Transport Medium ◽  
Viral Transport ◽  
Viral Transport Medium ◽  
Stool Specimens

Two media for preserving the viability of different viruses in clinical material during transit at ambient temperature have been developed. One was designed for transporting stool specimens and autopsy material, while the other was designed for such specimens as throat washings, swabs, and cerebrospinal fluid. The essential ingredient in both media was bentonite either uncoated or coated with serum proteins. Preparation of both media is given in detail.Experimental results presented indicate that coxsackie A9, B5, echo 11, adeno 5, influenza A2, parainfluenza, rubella, and herpes simplex viruses could sustain their infectivity without loss of titer in bentonite transport medium for prolonged time, from 3 to 21 days, depending on the virus type.The bentonite transport medium was found to be superior to commonly used charcoal viral transport medium.

Comparison of nasopharyngeal nylon flocked swabs with universal transport medium and rayon-bud swabs with a sponge reservoir of viral transport medium in the diagnosis of paediatric influenza

2010 ◽  
Vol 59 (1) ◽  
pp. 96-99 ◽  
Author(s):  
Susanna Esposito ◽  
Claudio Giuseppe Molteni ◽  
Cristina Daleno ◽  
Antonia Valzano ◽  
Laura Cesati ◽  
...  
Keyword(s):  
Influenza A ◽  
Molecular Detection ◽  
Influenza Viruses ◽  
Influenza B ◽  
Threshold Values ◽  
Detection Rates ◽  
Laboratory Staff ◽  
Transport Medium ◽  
Viral Transport ◽  
Viral Transport Medium

This study compared a kit containing a nasopharyngeal nylon flocked swab and a tube with a liquid universal transport medium (UTM) with a kit containing a plastic-shafted rayon-budded swab with a sponge reservoir of viral transport medium for the molecular detection of influenza viruses in children. Respiratory samples were collected from 314 children aged <5 years with influenza-like illness (186 males; mean age 2.32±2.27 years) using both swabs in a randomized sequence for each patient. The flocked swabs permitted the detection of 28 influenza A (8.9 %) and 45 influenza B (14.3 %) cases, and the rayon-bud swabs 26 influenza A (8.3 %) and 43 influenza B (13.7 %) cases, with detection rates of 23.2 and 22.0 %, respectively, and similar cycle threshold values. Paediatricians and laboratory staff were significantly more satisfied with both the simplicity (P <0.0001) and rapidity (P <0.0001) of the nasopharyngeal flocked swabs with UTM. These findings show that the flocked swabs with UTM and the rayon-bud swabs with a sponge transport medium are similarly efficient in preserving influenza virus nucleic acid, but that the kit containing a flocked swab with a UTM allows easier and more rapid collection and processing of specimens.

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.

Sign in / Sign up

Export Citation Format

Share Document