scholarly journals Viral Load Dynamics in Sputum and Nasopharyngeal Swab in Patients with COVID-19

2020 ◽  
Vol 99 (11) ◽  
pp. 1239-1244 ◽  
Author(s):  
R. Liu ◽  
S. Yi ◽  
J. Zhang ◽  
Z. Lv ◽  
C. Zhu ◽  
...  
Keyword(s):  
Viral Load ◽  
Quantitative Polymerase Chain Reaction ◽  
Nasopharyngeal Swab ◽  
Chain Reaction ◽  
Global Pandemic ◽  
Observational Cohort ◽  
Polymerase Chain ◽  
Underlying Diseases ◽  
Temporal Profiles ◽  
Substantial Morbidity

Coronavirus disease 2019 (COVID-19) has caused a global pandemic associated with substantial morbidity and mortality. Nasopharyngeal swabs and sputum samples are generally collected for serial viral load screening of respiratory contagions, but temporal profiles of these samples are not completely clear in patients with COVID-19. We performed an observational cohort study at Renmin Hospital of Wuhan University, which involved 31 patients with confirmed COVID-19 with or without underlying diseases. We obtained samples from each patient, and serial viral load was measured by real-time quantitative polymerase chain reaction. We found that the viral load in the sputum was inclined to be higher than samples obtained from the nasopharyngeal swab at disease presentation. Moreover, the viral load in the sputum decreased more slowly over time than in the nasopharyngeal group as the disease progressed. Interestingly, even when samples in the nasopharyngeal swab turned negative, it was commonly observed that patients with underlying diseases, especially hypertension and diabetes, remained positive for COVID-19 and required a longer period for the sputum samples to turn negative. These combined findings emphasize the importance of tracking sputum samples even in patients with negative tests from nasopharyngeal swabs, especially for those with underlying conditions. In conclusion, this work reinforces the importance of sputum samples for SARS-CoV-2 detection to minimize transmission of COVID-19 within the community.

scholarly journals Detection of HBV DNA by PCR and its application in clinical transfusion

2018 ◽  
Vol 1 (1) ◽  
pp. 22
Author(s):  
Shunqing Li
Keyword(s):  
Viral Load ◽  
Hepatitis B ◽  
Quantitative Polymerase Chain Reaction ◽  
Hbv Dna ◽  
Serological Markers ◽  
Serologic Test ◽  
Chain Reaction ◽  
Medical Disputes ◽  
B Virus ◽  
Polymerase Chain

<p><strong><em> </em></strong>This study was to detect the hepatitis B virus (HBV) DNA copies in patients through blood transfusions; recessive carriers with HBsAg negative but HBV DNA positive were further studied to see the content and distribution of HBV in patients, and provide evidence for the clinical treatment. A total of 532 blood samples collected from July 2014 to July 2015 were tested for HBV-DNA viral load and hepatitis B serological markers using quantitative Polymerase Chain Reaction (qPCR) and serologic test (five serological markers of hepatitis B). The results showed that, 3 cases were HBV serology negative and the HBV-DNA viral load was in the range of 250-500 whereas only 1 case was HBsAb positive and the HBV-DNA viral load was above 500. qPCR, for detecting HBV DNA, together with serological routine test can effectively reduce HBV infection during transfusion and prevent medical disputes.</p>

scholarly journals Detection of HBV DNA by PCR and its application in clinical transfusion

2018 ◽  
Vol 1 (1) ◽  
pp. 22
Author(s):  
Shunqing Li
Keyword(s):  
Viral Load ◽  
Hepatitis B ◽  
Quantitative Polymerase Chain Reaction ◽  
Hbv Dna ◽  
Serological Markers ◽  
Serologic Test ◽  
Chain Reaction ◽  
Medical Disputes ◽  
B Virus ◽  
Polymerase Chain

<p><strong><em> </em></strong>This study was to detect the hepatitis B virus (HBV) DNA copies in patients through blood transfusions; recessive carriers with HBsAg negative but HBV DNA positive were further studied to see the content and distribution of HBV in patients, and provide evidence for the clinical treatment. A total of 532 blood samples collected from July 2014 to July 2015 were tested for HBV-DNA viral load and hepatitis B serological markers using quantitative Polymerase Chain Reaction (qPCR) and serologic test (five serological markers of hepatitis B). The results showed that, 3 cases were HBV serology negative and the HBV-DNA viral load was in the range of 250-500 whereas only 1 case was HBsAb positive and the HBV-DNA viral load was above 500. qPCR, for detecting HBV DNA, together with serological routine test can effectively reduce HBV infection during transfusion and prevent medical disputes.</p>

scholarly journals APLIKASI KUANTIFIKASI KOI HERPESVIRUS : REAL TIME – QUANTITATIVE POLYMERASE CHAIN REACTION (RT-Q PCR) MENGGUNAKAN SYBR GREEN PADA IKAN MAS (Cyprinus carpio)

2017 ◽  
Vol 12 (1) ◽  
pp. 45
Author(s):  
Isti Koesharyani ◽  
Lila Gardenia ◽  
Tatik Mufidah ◽  
Ayi Santka
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Load ◽  
Real Time ◽  
Cyprinus Carpio ◽  
Quantitative Polymerase Chain Reaction ◽  
Sybr Green ◽  
Chain Reaction ◽  
Koi Herpesvirus ◽  
Ct Value ◽  
Polymerase Chain

Koi Herpes Virus (KHV) di Indonesia sejak tahun 2002 merupakan penyakit mematikan yang menyerang ikan koi Cyprinus carpio koi dan ikan mas Cyprinus carpio carpio, dan sampai saat ini, infeksi KHV dilaporkan sudah menyebar hampir di seluruh dunia. Untuk mengetahui adanya infeksi KHV perlu cara diagnosa yang sangat akurat/sensitif, sehingga keberadaan KHV dapat diketahui secara pasti dengan tingkat sensitivitas yang lebih baik pada ikan budidaya. Tujuan dari penelitian ini adalah untuk mengaplikasikan teknik deteksi dengan real time quantitative polymerase chain reaction (RT- qPCR/qPCR) guna mengetahui adanya infeksi KHV secara kuantitatif pada ikan mas dengan mengetahui kandungan virus (viral load). Sebanyak masing-masing 3 ekor sampel diperoleh dari sentra budidaya ikan mas di Cirata-Jawa Barat, Maninjau-Sumatera Barat, dan Banjarmasin-Kalimantan Selatan. Sampel-sampel tersebut selanjutnya dianalisa keberadaan KHV-nya dengan RT-qPCR menggunakan SYBR Green. Hasil pengujian menunjukkan bahwa jumlah tertinggi (viral load) diperoleh dari ikan mas asal Cirata-3 dengan nilai Threshold Cycle (Ct.) 18,24 atau setara dengan 3,4 x 107 kopi, dan terendah dari ikan mas asal Banjarmasin-3 dengan nilai Ct. 33,39 atau 1,8 x 102 kopi. Dua standar yang digunakan dalam pengujian ini berupa plasmid dengan jumlah kopi 2 x 104 (Ct 27,24) dan 2 x 103 (Ct 30,24) dan kontrol atau Non Template Control (NTC) adalah 3,1 x 10 atau dengan nilai Ct 35,65. Uji aplikasi deteksi KHV dengan metode RT-qPCR ini memberikan hasil yang lebih sensitif, di mana sampel yang tidak terdeteksi dengan metode PCR konvensional dapat dideteksi dan dihitung jumlah kopi DNA (DNA copy). Since 2002, Koi herpesvirus (KHV) in Indonesian has been a malignant diseases, now recognized as a worldwide cause of mortality among populations of koi Cyprinus carpio koi and common carp Cyprinus carpio carpio. To determine the presence of infection is required the KHV diagnosis method with highly accurate and sensitive, so that the existence KHV can be known exactly with high sensitivity level in fish farming.The objective of this study was to develop and evaluate the infection by Real Time Quantitative Polymerase Chain Reaction (RT- qPCR/qPCR). Sample were taken from Carp culture in West Java, West Sumatra, and South Kalimantan. The assay was done by SYBR Green RT-qPCR. The analysis result of KHV in carp revealed that the carp from Cirata-3 had the highest viral load with Ct. value 18.24 equal with 3.4 x 107 copies, and the lowest one was the carp from Banjarmasin-3 at Ct. value 33.39 (1.8 x 102 copies), while two standards plasmid and Non Template Control (NTC) had Ct value of 27.24 (2 x 104copies),30.24 (2 x 103copies), and 35.65 (3.1 x 10 copies), respectively. Application KHV test by q-PCR has more advantages and sensitive than that of conventional PCR, and it can be used to detect and calculate the copy number of DNA.

scholarly journals The handling of SARS-CoV-2 associated deaths - infectivity of the body

2021 ◽  
Author(s):  
Ann Sophie Schröder ◽  
Carolin Edler ◽  
Benjamin Ondruschka ◽  
Klaus Püschel ◽  
Julia Schädler ◽  
...  
Keyword(s):  
Body Surface ◽  
Quantitative Polymerase Chain Reaction ◽  
The Body ◽  
Nasopharyngeal Swab ◽  
Post Mortem ◽  
Chain Reaction ◽  
Body Regions ◽  
Occupational Infection ◽  
Polymerase Chain ◽  
Adequate Management

AbstractThe body of a deceased with Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection is considered infectious. In this study, we present the results of infectivity testing of the body and testing of mortuary staff for SARS-CoV-2. We performed real-time quantitative polymerase chain reaction (RT-qPCR) for SARS-CoV-2 on 33 decedents with ante mortem confirmed SARS-CoV-2 infection. Swabs of the body surface from five different body regions and from the body bag or coffin were examined. A subset of the swabs was brought into cell culture. In addition, screening of 25 Institute of Legal Medicine (ILM) personnel for ongoing or past SARS-CoV-2 infection was performed at two different time points during the pandemic. Swabs from all locations of the body surface and the body environment were negative in cases of negative post mortem nasopharyngeal testing (n=9). When the post mortem nasopharyngeal swab tested positive (n=24), between 0 and 5 of the body surface swabs were also positive, primarily the perioral region. In six of the cases, the body bag also yielded a positive result. The longest postmortem interval with positive SARS-CoV-2 RT-qPCR at the body surface was nine days. In no case viable SARS-CoV-2 was found on the skin of the bodies or the body bags. One employee (autopsy technician) had possible occupational infection with SARS-CoV-2; all other employees were tested negative for SARS-CoV-2 RNA or antibody twice. Our data indicate that with adequate management of general safety precautions, transmission of SARS-CoV-2 through autopsies and handling of bodies is unlikely.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

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