scholarly journals A Study on Requirement of Cold Chain Maintenance for Reliable Testing of SARS-CoV-2 Samples

2021 ◽  
Author(s):  
Varunika Vijayvergia ◽  
Aruna Vyas ◽  
Nazneen Pathan ◽  
Rajni Sharma ◽  
Snigdha Purohit ◽  
...  
Keyword(s):  
Room Temperature ◽  
Standard Test ◽  
Cold Chain ◽  
Effect Of Temperature ◽  
Sample Collection ◽  
Rt Pcr ◽  
Significant Deterioration ◽  
Clinical Sensitivity ◽  
Ambient Room Temperature ◽  
The World

Introduction: Coronavirus Disease 2019 (COVID-19) has been haunting the world since December 2019 and has grown to pandemic proportions from March 2020. Even after a full year of research and study, the most effective way to control the spread of this infection is early diagnosis and isolation of the cases. Real-time Reverse Transcription Polymerase Chain Reaction (RT-PCR) is considered the standard test all over the world for the diagnosis of Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) infection. All the sample collection guidelines have recommended stringent maintenance of the cold chain for the sample transport. However, it is not possible for the resource constrained developing countries with inadequate infrastructure to comply with these guidelines all the time. Aim: To determine necessity of stringent transport criteria and the effect of temperature on the clinical sensitivity of a RT-PCR assay for diagnosis of SARS-CoV-2 infection. Materials and Methods: In this prospective experimental study conducted in November 2020, 49 positive samples were kept at ambient room temperature and were tested everyday with RT- PCR for the detection of SARS-CoV-2 Ribonucleic Acid (RNA). The samples were also kept under refrigeration at 4°C and were also tested by RT-PCR and the results were compared with their respective counterparts kept at room temperature till nine days. Python Jupiter notebook SciPy and Anaconda software was used for statistical analysis. Results: It was observed that the positivity of the RT-PCR results were not deteriorated till five days and there was no significant deterioration even after nine days of samples being stored at room temperature suggesting that even if the viral RNA itself is not stable outside strict temperature control but small fragment or target genetic sequences are enough for detection of virus by RT-PCR. Conclusion: It is possible to keep samples at this ambient temperature for five days without any loss of positivity in RT-PCR.

scholarly journals Self-collected gargle specimen as a patient-friendly sample collection method for COVID-19 diagnosis in a population context

2021 ◽  
Author(s):  
Revata Utama ◽  
Rebriarina Hapsari ◽  
Iva Puspitasari ◽  
Desvita Sari ◽  
Meita Hendrianingtyas ◽  
...  
Keyword(s):  
Healthcare Workers ◽  
Room Temperature ◽  
Rna Extraction ◽  
User Preference ◽  
Sample Collection ◽  
Rt Pcr ◽  
Collection Method ◽  
Qrt Pcr ◽  
Alternative Specimen ◽  
Temperature Storage

Abstract Scaling up SARS-CoV-2 testing and tracing continues to be plagued with the limitation of the sample collection method, which requires trained healthcare workers to perform and causes discomfort to the patients. In response, we assessed the performance and user preference of gargle specimens for qRT-PCR-based detection of SARS-CoV-2 in Indonesia. Inpatients who had recently been diagnosed with COVID-19 and outpatients who were about to perform qRT-PCR testing were asked to provide nasopharyngeal and oropharyngeal (NPOP) swabs and self-collected gargle specimens. We demonstrated that self-collected gargle specimens can be an alternative specimen to detect SARS-CoV-2 and the viral RNA remained stable for 31 days at room temperature storage. The developed method was validated for use on multiple RNA extraction kits and commercially available COVID-19 RT-PCR kits. Our developed method achieved a sensitivity of 91.38% when compared to paired NPOP swab specimens (Ct < 35), with 97.10% of patients preferring the self-collected gargle method.

scholarly journals Self-collected gargle specimen as a patient-friendly sample collection method for COVID-19 diagnosis in a population context

2021 ◽  
Author(s):  
Revata Utama ◽  
Rebriarina Hapsari ◽  
Iva Puspitasari ◽  
Desvita Sari ◽  
Meita Hendrianingtyas ◽  
...  
Keyword(s):  
Healthcare Workers ◽  
Room Temperature ◽  
Rna Extraction ◽  
User Preference ◽  
Sample Collection ◽  
Rt Pcr ◽  
Collection Method ◽  
Qrt Pcr ◽  
Alternative Specimen ◽  
Temperature Storage

Abstract Scaling up SARS-CoV-2 testing and tracing continues to be plagued with the limitation of the sample collection method, which requires trained healthcare workers to perform and causes discomfort to the patients. In response, we assessed the performance and user preference of gargle specimens for qRT-PCR-based detection of SARS-CoV-2 in Indonesia. Inpatients who had recently been diagnosed with COVID-19 and outpatients who were about to perform qRT-PCR testing were asked to provide nasopharyngeal and oropharyngeal (NPOP) swabs and self-collected gargle specimens. We demonstrated that self-collected gargle specimens can be an alternative specimen to detect SARS-CoV-2 and the viral RNA remained stable for 31 days at room temperature storage. The developed method was validated for use on multiple RNA extraction kits and commercially available COVID-19 RT-PCR kits. Our developed method achieved a sensitivity of 91.38% when compared to paired NPOP swab specimens (Ct < 35), with 97.10% of patients preferring the self-collected gargle method.

Self-collected gargle as a patient-friendly sample collection method for COVID-19 diagnosis in population context

2021 ◽  
Author(s):  
Revata Utama ◽  
Rebriarina Hapsari ◽  
Iva Puspitasari ◽  
Desvita Sari ◽  
Meita Hendrianingtyas ◽  
...  
Keyword(s):  
Healthcare Workers ◽  
Room Temperature ◽  
Rna Extraction ◽  
User Preference ◽  
Sample Collection ◽  
Rt Pcr ◽  
Collection Method ◽  
Qrt Pcr ◽  
Alternative Specimen ◽  
Temperature Storage

Abstract Scaling up SARS-CoV-2 testing and tracing continues to be plagued with limitation of sample collection method that requires trained healthcare workers to perform and cause discomfort to the patients. In response, we assessed the performance and user preference of gargle specimens for qRT-PCR based detection of SARS-CoV-2 in Indonesia. Inpatients who had recently been diagnosed with COVID-19 and outpatients who were about to perform qRT-PCR testing were asked to provide nasopharyngeal and oropharyngeal (NPOP) swabs and self-collected gargle specimens. We demonstrated that self-collected gargle specimens can be an alternative specimen to detect SARS-CoV-2 and the viral RNA remained stable for 31 days on room temperature storage. The developed method was validated for use on multiple RNA extraction kit and commercially available COVID-19 RT-PCR kits. Our developed method achieved sensitivity of 91.38% when compared to paired NPOP swab specimens (Ct < 35) with 95.16% of patients prefer the self-collected gargle method.

Growth of Salmonella enterica and Listeria monocytogenes on Fresh-Cut Cantaloupe under Different Temperature Abuse Scenarios

2015 ◽  
Vol 78 (6) ◽  
pp. 1125-1131 ◽  
Author(s):  
JINGWEI HUANG ◽  
YAGUANG LUO ◽  
XIANGWU NOU
Keyword(s):  
Listeria Monocytogenes ◽  
Salmonella Enterica ◽  
Room Temperature ◽  
Storage Temperature ◽  
Cold Chain ◽  
Significant Deterioration ◽  
Tissue Integrity ◽  
Cell Counts ◽  
Fresh Cut ◽  
The Impact

Effective cold chain management is a critical component of food safety practice. In this study, we examined the impact of commonly encountered temperature abuse scenarios on the proliferation of Salmonella enterica and Listeria monocytogenes on fresh-cut cantaloupe. Inoculated fresh-cut cantaloupe cubes were subjected to various temperature abuse conditions, and the growth of S. enterica and L. monocytogenes was determined. During 1 week of storage, Salmonella cell counts on fresh-cut cantaloupe increased by −0.26, 1.39, and 2.23 log units at 4°C (control), 8°C, and 12°C (chronic temperature abuse), respectively, whereas that of L. monocytogenes increased by 0.75, 2.86, and 4.17 log units. Under intermittent temperature abuse conditions, where storage temperature fluctuated twice daily to room temperature for 30 min, Salmonella cell count increased by 2.18 log units, whereas that of L. monocytogenes increased by 1.86 log units. In contrast, terminal acute temperature abuses for 2 to 4 h resulted in upwards to 0.6 log unit for Salmonella, whereas the effect on L. monocytogenes was less significant compared with L. monocytogenes on cut cantaloupe stored at 4°C. Significant deterioration of produce visual quality and tissue integrity, as reflected by electrolyte leakage, was also observed under various temperature abuse conditions.

Addressing the constraints of Tritrichomonas foetus sample collection in remote areas: lyophilized modified Diamond's media as a substitute for liquid medium

Parasitology ◽  
2019 ◽  
Vol 146 (9) ◽  
pp. 1184-1187 ◽  
Author(s):  
Gemma Rush ◽  
Michael William Reynolds ◽  
Nichola Eliza Davies Calvani ◽  
Jan Šlapeta
Keyword(s):  
Liquid Medium ◽  
Room Temperature ◽  
Cold Chain ◽  
Tritrichomonas Foetus ◽  
Robust Method ◽  
Sample Collection ◽  
Remote Areas ◽  
Reproductive Disease ◽  
The Media

AbstractBovine trichomoniasis is a notifiable, reproductive disease of cattle caused by the parasite Tritrichomonas foetus. Culturing with modified Diamond's medium (MDM) is required to increase the low number of organisms received from a preputial sample, but is limited in application to remote areas as it requires continuous cold chain storage. This study utilized lyophilization to sustain the viability of MDM during transport in lieu of a continuous cold chain. All lyophilized MDM was able to sustain T. foetus after storage for 42 days at 24 °C, and the results demonstrated that lyophilized MDM was equally as viable as refrigerated liquid MDM. Storage of lyophilized MDM at room temperature for 1 and 7 days did not impact T. foetus yield, both with and without exposure to light. A limitation of the lyophilized MDM was demonstrated with a significant decrease in T. foetus yield when the media was stored at 37 and 58 °C. The lyophilization of MDM provides a robust method of transporting and storing medium prior to reconstitution and inoculation, for use in T. foetus diagnosis and surveillance in remote areas.

scholarly journals Saliva Is Comparable to Nasopharyngeal Swabs for Molecular Detection of SARS-CoV-2

2021 ◽  
Author(s):  
Cody Callahan ◽  
Sarah Ditelberg ◽  
Sanjucta Dutta ◽  
Nancy Littlehale ◽  
Annie Cheng ◽  
...  
Keyword(s):  
High Sensitivity ◽  
Outpatient Setting ◽  
Viral Rna ◽  
Molecular Testing ◽  
Sample Collection ◽  
Rt Pcr ◽  
Clinical Sensitivity ◽  
Viral Loads ◽  
Infectious Risk ◽  
A Minor

AbstractBackgroundThe continued need for molecular testing for SARS-CoV-2 and potential for self-collected saliva as an alternative to nasopharyngeal (NP) swabs for sample acquisition led us to compare saliva to NP swabs in an outpatient setting, without restrictions to avoid food, drink, smoking, or tooth-brushing.MethodsA total of 385 pairs of NP and saliva specimens were obtained, the majority from individuals presenting for initial evaluation, and were tested on two high-sensitivity RT-PCR platforms: the Abbott m2000 and Abbott Alinity m (both with limits of detection [LoD] of 100 copies of viral RNA/mL).ResultsConcordance between saliva and NP was excellent overall (Cohen’s κ=0.93), for both initial and followup testing, for both platforms, and for specimens treated with guanidinium transport medium as preservative as well as for untreated saliva (κ=0.88-0.95). Viral loads were on average 16x higher in NP specimens than saliva specimens, suggesting that only the relatively small fraction of outpatients (∼8% in this study) who present with very low viral loads (<1,600 copies/mL from NP swabs) would be missed by testing saliva instead of NP swabs, when using sensitive testing platforms. Special attention was necessary to ensure leak-resistant specimen collection and transport.ConclusionsThe advantages of self-collection of saliva, without behavioral restrictions, will likely outweigh a minor potential decrease in clinical sensitivity in individuals less likely to pose an infectious risk to others for many real-world scenarios, especially for initial testing.Key pointsSaliva has comparable sensitivity and specificity to nasopharyngeal swabs for RT-PCR-based COVID-19 testing (concordance, κ=0.93; n=385 participants), albeit with slightly lower recovery of viral RNA. Treatment with a readily available guanidinium preservative within 24 hours of sample collection improves recovery.

Effect of Temperature on ADP-Induced Platelet Aggregation

1973 ◽  
Vol 29 (01) ◽  
pp. 183-189
Author(s):  
C. A Praga ◽  
E. M Pogliani
Keyword(s):  
Platelet Aggregation ◽  
Incubation Period ◽  
Room Temperature ◽  
Important Variable ◽  
Practical Significance ◽  
Effect Of Temperature ◽  
Induce Platelet Aggregation ◽  
Cuvette Holder ◽  
Exact Temperature

SummaryTemperature represents a very important variable in ADP-induced platelet aggregation.When low doses of ADP ( < 1 (μM) are used to induce platelet aggregation, the length of the incubation period of PRP in the cuvette holder of the aggregometer, thermostatted at 37° C, is very critical. Samples of the same PRP previously kept at room temperature, were incubated for increasing periods of time in the cuvette of the aggregometer before adding ADP, and a significant decrease of aggregation, proportional to the length of incubation, was observed. Stirring of the PRP during the incubation period made these changes more evident.To measure the exact temperature of the PRP during incubation in the aggre- gometer, a thermocouple device was used. While the temperature of the cuvette holder was stable at 37° C, the PRP temperature itself increased exponentially, taking about ten minutes from the beginning of the incubation to reach the value of 37° C. The above results have a practical significance in the reproducibility of the platelet aggregation test in vitro and acquire particular value when the effect of inhibitors of ADP induced platelet aggregation is studied.Experiments carried out with three anti-aggregating agents (acetyl salicyclic acid, dipyridamole and metergoline) have shown that the incubation conditions which influence both the effect of the drugs on platelets and the ADP breakdown in plasma must be strictly controlled.

EFEKTIVITAS UJIAN AKHIR SEMESTER SECARA ONLINE MENGGUNAKAN APLIKASI GOOGLE FORM PADA SMP MA'ARIF NU CIMANGGU

2021 ◽  
Vol 2 (2) ◽  
pp. 76-86
Author(s):  
Diky Setiawan ◽  
Mochammad Hasymi Somaida
Keyword(s):  
Survey Method ◽  
Sample Collection ◽  
Quantitative Survey ◽  
Corona Virus ◽  
The World ◽  
Teachers And Students ◽  
The Impact

The Corona virus is a virus that originates from China, and spreads rapidly throughout the world andbegan to spread to Indonesia in early March 2020. The impact of Covid-19 has caused losses in all fields,be it economy, education, etc. In the field of education, activities both learning and examinations must bereplaced online due to the impact of the spread of Covid-19. This study aims to determine theeffectiveness of the implementation of UAS which is carried out online using the Google Form applicationmedia at SMP MA'ARIF NU CIMANGGU. This research uses the quantitative survey method whichdescribes the online UAS implementation activities at SMP MA'ARIF NU CIMANGGU during thepandemic. The object consists of 35 student respondents. The data sample collection is done using aquestionnaire / questionnaire containing questions related to the implementation of UAS online using theGoogle Form application at SMP MA ' ARIF NU CIMANGGU. Based on the results of this study, theimplementation of online UAS runs effectively f and good, it can be seen from the results of satisfactorystudent scores. The application used is of course the Googke Form as the main media, and Whatsapp as amedium for interaction between teachers and students. The obstacles experienced are regarding badconnections, limited quota, schedules that frequently change, as well as difficulties in understandingsubject matter.Keywords: Covid-19,impact,effectiveness

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 Green Biosynthesis of Silver Nanoparticles Using Leaf Extract of Carissa carandas L. and Their Antioxidant and Antimicrobial Activity against Human Pathogenic Bacteria

Biomolecules ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 299
Author(s):  
Reetika Singh ◽  
Christophe Hano ◽  
Gopal Nath ◽  
Bechan Sharma
Keyword(s):  
Silver Nanoparticles ◽  
Pathogenic Bacteria ◽  
Leaf Extract ◽  
Room Temperature ◽  
Xrd Analysis ◽  
Antibacterial Activities ◽  
Effect Of Temperature ◽  
Human Pathogenic Bacteria ◽  
Carissa Carandas ◽  
Antioxidant And Antimicrobial Activity

Carissa carandas L. is traditionally used as antibacterial medicine and accumulates many antioxidant phytochemicals. Here, we expand this traditional usage with the green biosynthesis of silver nanoparticles (AgNPs) achieved using a Carissa carandas L. leaf extract as a reducing and capping agent. The green synthesis of AgNPs reaction was carried out using 1mM silver nitrate and leaf extract. The effect of temperature on the synthesis of AgNPs was examined using room temperature (25 °C) and 60 °C. The silver nanoparticles were formed in one hour by stirring at room temperature. In this case, a yellowish brown colour was developed. The successful formation of silver nanoparticles was confirmed by UV–Vis, Fourier transform infrared (FT-IR) and X-ray diffraction (XRD) analysis. The characteristic peaks of the UV-vis spectrum and XRD confirmed the synthesis of AgNPs. The biosynthesised AgNPs showed potential antioxidant activity through DPPH assay. These AgNPs also exhibited potential antibacterial activity against human pathogenic bacteria. The results were compared with the antioxidant and antibacterial activities of the plant extract, and clearly suggest that the green biosynthesized AgNPs can constitute an effective antioxidant and antibacterial agent.

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