scholarly journals Comparison of Rates of Positivity for Bordetella pertussis by Real-Time PCR between Specimens Collected with Rayon Swabs on Aluminum Wire Shaft in Amies Gel with Charcoal and Specimens Collected with Flocked Swabs in Universal Viral Transport Medium during an Epidemic

2014 ◽  
Vol 52 (7) ◽  
pp. 2656-2658 ◽  
Author(s):  
S. Arbefeville ◽  
P. Ferrieri
Keyword(s):  
Real Time ◽  
Bordetella Pertussis ◽  
Real Time Pcr ◽  
Aluminum Wire ◽  
Transport Medium ◽  
Viral Transport ◽  
Viral Transport Medium

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 Addition of a Homologous Internal Control to a Real-Time PCR Assay for Detection of Bordetella pertussis

2005 ◽  
Vol 51 (12) ◽  
pp. 2404-2406 ◽  
Author(s):  
Christoph Koidl ◽  
Michael Bozic ◽  
Jörg Berg ◽  
Markus Stöcher ◽  
Gerhard Mühlbauer ◽  
...  
Keyword(s):  
Real Time ◽  
Bordetella Pertussis ◽  
Internal Control ◽  
Real Time Pcr ◽  
Pcr Assay

scholarly journals Evaluation of Real-Time PCR for Detection of and Discrimination between Bordetella pertussis, Bordetella parapertussis, and Bordetella holmesii for Clinical Diagnosis

2003 ◽  
Vol 41 (9) ◽  
pp. 4121-4126 ◽  
Author(s):  
K. E. Templeton ◽  
S. A. Scheltinga ◽  
A. van der Zee ◽  
B. M. W. Diederen ◽  
A. M. Kruijssen ◽  
...  
Keyword(s):  
Real Time ◽  
Clinical Diagnosis ◽  
Bordetella Pertussis ◽  
Real Time Pcr ◽  
Bordetella Parapertussis

scholarly journals Comparison of Serological and Real-Time PCR Assays To Diagnose Bordetella pertussis Infection in 2007

2008 ◽  
Vol 46 (5) ◽  
pp. 1672-1677 ◽  
Author(s):  
P. Andre ◽  
V. Caro ◽  
E. Njamkepo ◽  
A. M. Wendelboe ◽  
A. Van Rie ◽  
...  
Keyword(s):  
Real Time ◽  
Bordetella Pertussis ◽  
Real Time Pcr ◽  
Pertussis Infection ◽  
Pcr Assays

Comparison of real-time PCR and pyrosequencing for typing Bordetella pertussis toxin subunit 1 variants

2006 ◽  
Vol 65 (1) ◽  
pp. 153-158 ◽  
Author(s):  
Martin Storm ◽  
Abdolreza Advani ◽  
Monica Pettersson ◽  
Hans O. Hallander ◽  
Kåre Bondeson
Keyword(s):  
Real Time ◽  
Pertussis Toxin ◽  
Bordetella Pertussis ◽  
Real Time Pcr ◽  
Bordetella Pertussis Toxin

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 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.

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