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  1. Article ; Online: Evaluation of RT-PCR assays for detection of SARS-CoV-2 variants of concern.

    Dip, Sourav Dutta / Sarkar, Shovon Lal / Setu, Md Ali Ahasan / Das, Prosanto Kumar / Pramanik, Md Hasan Ali / Alam, A S M Rubayet Ul / Al-Emran, Hassan M / Hossain, M Anwar / Jahid, Iqbal Kabir

    Scientific reports

    2023  Volume 13, Issue 1, Page(s) 2342

    Abstract: Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) pandemic has been considered with great importance on correct screening procedure. The detection efficiency of recent variants of concern were observed by comparing 5 commercial RT-PCR kits and ...

    Abstract Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) pandemic has been considered with great importance on correct screening procedure. The detection efficiency of recent variants of concern were observed by comparing 5 commercial RT-PCR kits and a SYBR-green method developed and validated in our laboratory. The RNA was extracted from nasopharyngeal samples from suspected COVID-19 patients and RT-PCR assay was performed according to the instruction of the respective manufacturers. The specificity and sensitivity of Maccura kit was 81.8% and 82.5%, A*Star kit was 100% and 75.4%, Da An Gene kit was 100% and 68.4%, Sansure kit was 54.5% and 91.2% and TaqPath kit was 100% and 70.2% respectively. Our in house SYBR-Green method showed a consistent detection result with 90.9% specificity and 91.2% sensitivity. We also found that detection kits targeting more genes showed better accuracy which facilitates less false positive results (< 20%). Our study found a significant difference (p < 0.005) in Ct value reported for common target genes shared by the RT-PCR kits in relation with different variants of COVID-19 infection. Recent variants of concerns contain more than 30 mutations in the spike proteins including 2 deletion and a unique insertion mutation by which makes detection of these variants difficult and these facilitates the variants to escape from being detected.
    MeSH term(s) Humans ; SARS-CoV-2/genetics ; COVID-19/diagnosis ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Viral/genetics ; RNA, Viral/analysis ; Sensitivity and Specificity ; COVID-19 Testing
    Chemical Substances RNA, Viral
    Language English
    Publishing date 2023-02-09
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-023-28275-y
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Relative expression of proinflammatory molecules in COVID-19 patients who manifested disease severities.

    Nigar, Shireen / Shah, Sm Tanjil / Setu, Md Ali Ahasan / Dip, Sourav Dutta / Ibnat, Habiba / Islam, M Touhidul / Akter, Selina / Jahid, Iqbal Kabir / Hossain, M Anwar

    Journal of medical virology

    2021  Volume 93, Issue 10, Page(s) 5805–5815

    Abstract: Aggressive immune response, due to overexpressed proinflammatory molecules, has been characterized in coronavirus disease 2019 (COVID-19) patients. Some of those mediators have a dual and opposite role on immune systems at play behind differential ... ...

    Abstract Aggressive immune response, due to overexpressed proinflammatory molecules, has been characterized in coronavirus disease 2019 (COVID-19) patients. Some of those mediators have a dual and opposite role on immune systems at play behind differential disease severities. We investigated the expression of some cytokines and chemokines in COVID-19 patients in Bangladesh. We diagnosed the patients by detecting severe acute respiratory syndrome coronavirus 2 RNA in nasal swab samples by the real-time RT-PCR method. Thirty adult patients were preselected based on their disease severities and grouped into mild, moderate, and severe cases. Nine healthy volunteers participated in this study as a control. Relative expression of nine cytokines/chemokine in total leukocytes was semi-quantified in SYBRgreen-based real-time quantitative reverse-transcriptase polymerase chain reaction. We performed statistical tests on transformed log data using SPSS 24.0. At the onset of symptoms (Day 1), angiotensin-converting enzyme 2 (ACE2) (p < 0.05) and interleukin (IL)-6 (p > 0.05) were upregulated in all COVID-19 groups, although the expression levels did not significantly correlate with disease severities. However, expressions of IL-6, monocyte chemotactic protein-1, macrophage inflammatory protein-1α, tumor necrosis factor-α (TNF-α), RANTES (regulated upon activation, normal T cell expressed and secreted), and ACE2, on Day 14, were positively correlated with disease severities. Relative viral load at Day 1 showed no significant correlation with cytokine expression but had a significant positive correlation with RANTES and ACE2 expression on Day 14 (p < 0.05). Male patients had a higher level of IL-6 than female patients on Day 1 (p < 0.05). All COVID-19 patients showed upregulated cytokines and chemokines on Day 14 compared to Day 1 except TNF-α. Female patients had a higher expression of ACE2 and IL-12 on Day 14. Upregulated cytokines/chemokines at the convalescent stage, especially IL-6, may help in targeting anticytokine therapy in post-COVID-19 patients' management.
    MeSH term(s) Adult ; Bangladesh/epidemiology ; COVID-19/diagnosis ; COVID-19/epidemiology ; COVID-19/immunology ; COVID-19/virology ; Chemokines/blood ; Cytokines/blood ; Female ; Humans ; Male ; Middle Aged ; SARS-CoV-2/isolation & purification ; Severity of Illness Index ; Viral Load
    Chemical Substances Chemokines ; Cytokines
    Language English
    Publishing date 2021-06-12
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 752392-0
    ISSN 1096-9071 ; 0146-6615
    ISSN (online) 1096-9071
    ISSN 0146-6615
    DOI 10.1002/jmv.27112
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Relative expression of pro-inflammatory molecules in COVID-19 patients manifested disease severities.

    Nigar, Shireen / Shah, SM Tanjil / Setu, Ali Ahasan / Dip, Sourav Dutta / Ibnat, Habiba / Islam, M. Touhid / Akter, Selina / Jahid, Iqbal Kabir / Hossain, Md. Anwar

    medRxiv

    Abstract: Aggressive immune response, due to over-expressed pro-inflammatory molecules, had been characterized in COVID-19 patients. Some of those mediators have a dual and opposite role on immune-systems to play behind differential disease severities. We ... ...

    Abstract Aggressive immune response, due to over-expressed pro-inflammatory molecules, had been characterized in COVID-19 patients. Some of those mediators have a dual and opposite role on immune-systems to play behind differential disease severities. We investigated the expression of some cytokines and chemokines in COVID-19 patients in Bangladesh. We diagnosed the patients by detecting SARS-CoV-2 RNA in nasal swab samples by the real-time RT-PCR method. Thirty adult patients were preselected based on their disease severities and grouped into mild, moderate, and severe cases. Nine healthy volunteers participated in this study as control. Relative expression of nine cytokines/chemokine in total leukocytes was semi-quantified in SYBRgreen-based qRT-PCR. We performed statistical tests on transformed log data using SPSS 24.0. At the onset of symptoms (day-1), ACE2 (P < 0.05) and IL-6 (P > 0.05) were up-regulated in all COVID-19 groups, although expression levels did not significantly correlate with disease severities. However, expression of IL-6, MCP-1, MIP-1α, TNF-α, RANTES, and ACE2, on day-14, were positively correlated with disease severities. Relative viral load at day-1 showed no significant correlation with cytokine expression but had a significant positive correlation with RANTES and ACE2 expression on day-14 (P < 0.05). Male patients had a higher level of IL-6 than female patients on day-1 (P < 0.05). All COVID-19 patients showed up-regulated cytokines and chemokines on the day-14 compared to day-1 except TNF-α. Female patients had higher expression of ACE2 and IL-12 on day-14. Up-regulated cytokines/chemokines at the convalescent stage, especially IL-6, may target anti-cytokine therapy in post-COVID-19 patients management.
    Keywords covid19
    Language English
    Publishing date 2021-04-07
    Publisher Cold Spring Harbor Laboratory Press
    Document type Article ; Online
    DOI 10.1101/2021.04.01.21254770
    Database COVID19

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