A Comparative Evaluation between Lateral Flow Antigen Assay (LFA) and RT PCR | Chapter 3 | Research Perspectives of Microbiology and Biotechnology Vol. 4

 

The Present study proposes to look for RT-PCR cycle threshold correlation with positive Rapid COVID-19 LFA results and the ability of LFAs to be used as a proxy, inexpensive marker of infectiousness in mass settings given the present global post-pandemic scenario. Lateral flow antigen assays (LFA) are a quick and affordable way to detect COVID-19 in healthcare settings with limited infrastructure. Rapid diagnostic tests are essential for early, timely case identification that allows for supportive care and patient isolation. In light of the COVID-19 pandemic, molecular testing is still an expensive endeavor. Moreover, residual RNA cannot be distinguished from replication-competent viruses using molecular techniques. The present study is a retrospective cross-sectional hospital-based study of 1.25 years duration (September 2020- January 2022). Rapid COVID-19 LFA results were compared with Real-time PCR for detection of SARS-CoV-2 in nasopharyngeal swabs. Two hundred rapid antigen-positive nasopharyngeal swabs obtained from COVID-19 suspects/contacts/preoperative/ screening patients were subjected to RT-PCR to study the correlation with cycle threshold (CT) values obtained for all the antigen-positive cases. 200 Rapid COVID-19 LFA-positive samples were analysed in the present study. Amidst the LFA-positive samples included in the study 187 (93.5%) were found to have concordant results when subjected to the gold-standard Real-time PCR. Discordant results were documented in 13 (6.5%) COVID-19 LFA-positive samples which were found to be negative by RT-PCR. The average Cycle threshold values were found to be 23.75 for the E gene, 25.36 for the N gene and 24.07 for RdRp gene. The average PCR Cycle threshold of LFA positive cases remained significantly undeterred (p<0.5) throughout the time period of the study stipulating the undaunted viral load across the different waves of the pandemic. Maximum association of LFA positivity with symptom manifestation was seen during the 1st wave of COVID-19 (September-December 2020 in India). The association of symptoms with LFA test positivity reduced to a significant extent during the 3rd wave of the pandemic in January 2022 (p<0.5) indicating the reduced clinical severity but not infectivity of the SARS-CoV-2 infection during the 3rd wave of the pandemic. When compared to RT-PCR, lateral flow assay-based diagnostic techniques are more economically and technically convenient while maintaining a high degree of concordance. These tests offer a clear benefit in terms of expediting patient triage and, consequently, improving patient management. RT-PCR being highly sensitive assay can aid diagnosis in patients with low viral load and seems superior than the LFA which requires a higher vial load for positivity. Considering the above factors SARS-CoV-2 LFAs find utility as proxy infectivity indicating modalities for rapid triaging and patient management.

 

 

Author(s) Details

Neetha S. Murthy

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

M.N. Sumana

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

A. Tejashree

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

Vidyavathi B. Chitharagi

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

Rashmi P. Mahale

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

Murubagal Raghavendra Rao

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

G.S. Sowmya

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

Ranjitha Shankare Gowda

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

R. Deepashree

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

S.R. Sujatha

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

 

Please see the link:- https://doi.org/10.9734/bpi/rpmab/v4/458

A Comparative Evaluation between Lateral Flow Antigen Assay (LFA) and RT PCR | Chapter 3 | Research Perspectives of Microbiology and Biotechnology Vol. 4

 

The Present study proposes to look for RT-PCR cycle threshold correlation with positive Rapid COVID-19 LFA results and the ability of LFAs to be used as a proxy, inexpensive marker of infectiousness in mass settings given the present global post-pandemic scenario. Lateral flow antigen assays (LFA) are a quick and affordable way to detect COVID-19 in healthcare settings with limited infrastructure. Rapid diagnostic tests are essential for early, timely case identification that allows for supportive care and patient isolation. In light of the COVID-19 pandemic, molecular testing is still an expensive endeavor. Moreover, residual RNA cannot be distinguished from replication-competent viruses using molecular techniques. The present study is a retrospective cross-sectional hospital-based study of 1.25 years duration (September 2020- January 2022). Rapid COVID-19 LFA results were compared with Real-time PCR for detection of SARS-CoV-2 in nasopharyngeal swabs. Two hundred rapid antigen-positive nasopharyngeal swabs obtained from COVID-19 suspects/contacts/preoperative/ screening patients were subjected to RT-PCR to study the correlation with cycle threshold (CT) values obtained for all the antigen-positive cases. 200 Rapid COVID-19 LFA-positive samples were analysed in the present study. Amidst the LFA-positive samples included in the study 187 (93.5%) were found to have concordant results when subjected to the gold-standard Real-time PCR. Discordant results were documented in 13 (6.5%) COVID-19 LFA-positive samples which were found to be negative by RT-PCR. The average Cycle threshold values were found to be 23.75 for the E gene, 25.36 for the N gene and 24.07 for RdRp gene. The average PCR Cycle threshold of LFA positive cases remained significantly undeterred (p<0.5) throughout the time period of the study stipulating the undaunted viral load across the different waves of the pandemic. Maximum association of LFA positivity with symptom manifestation was seen during the 1st wave of COVID-19 (September-December 2020 in India). The association of symptoms with LFA test positivity reduced to a significant extent during the 3rd wave of the pandemic in January 2022 (p<0.5) indicating the reduced clinical severity but not infectivity of the SARS-CoV-2 infection during the 3rd wave of the pandemic. When compared to RT-PCR, lateral flow assay-based diagnostic techniques are more economically and technically convenient while maintaining a high degree of concordance. These tests offer a clear benefit in terms of expediting patient triage and, consequently, improving patient management. RT-PCR being highly sensitive assay can aid diagnosis in patients with low viral load and seems superior than the LFA which requires a higher vial load for positivity. Considering the above factors SARS-CoV-2 LFAs find utility as proxy infectivity indicating modalities for rapid triaging and patient management.

 

 

Author(s) Details

Neetha S. Murthy

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

M.N. Sumana

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

A. Tejashree

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

Vidyavathi B. Chitharagi

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

Rashmi P. Mahale

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

Murubagal Raghavendra Rao

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

G.S. Sowmya

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

Ranjitha Shankare Gowda

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

R. Deepashree

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

S.R. Sujatha

Department of Microbiology, JSS Academy of Higher Education and Research, Mysore, Karnataka, India.

 

Please see the link:- https://doi.org/10.9734/bpi/rpmab/v4/458

Clinical Efficacy and Safety of Remdesivir in Moderate to Severe COVID- 19 Disease in Patients Hospitalized in ICU | Chapter 4| Medicine and Medical Research: New Perspectives Vol. 8

Background: The COVID-19 pandemic has had a profound impact on global health and economies, and continues to pose significant challenges worldwide. Among the various drugs approved for treating COVID-19, Remdesivir, a purine nucleoside analogue inhibiting RNA-dependent RNA polymerase, has emerged as a key option in managing moderate-to-severe cases. This study evaluated the clinical efficacy and safety of Remdesivir in hospitalized patients with RT-PCR-confirmed moderate-to-severe COVID-19 disease. Conducted in the ICU of a hospital in Bihar, it provided real-world data on Remdesivir's impact on clinical outcomes, including hospital stay and mortality, particularly among patients with comorbidities. The study shows significant improvements in inflammatory markers post-treatment, highlighting the drug's role in managing the disease. It also adds valuable local data to the global understanding of COVID-19 treatment, guiding healthcare providers in similar settings.

Materials and Methods: This was a prospective, observational, cross-sectional study based on data collected from patients. A total of 100 patients with moderate-to-severe COVID-19 (RT-PCR positive) were admitted to the intensive care unit (ICU) of Sri Krishna Medical College and Hospital (SKMCH), Muzaffarpur, Bihar, India were enrolled in the study from 12th August to 11th November 2020. All patients with moderate to severe COVID-19 received an injection of Remdesivir, starting with a 200 mg intravenous (IV) loading dose on Day 1, followed by 100 mg IV daily for the next four days, in conjunction with other standard treatments.

Results: Out of 100 patients, 88 patients recovered and were discharged, while 12 patients died. The mean age of patients was 54 ± 16 years with male preponderance (4:1). Mean duration of hospital stay was 10.6 ± 5.4 days. C-reactive protein, D-dimer, ferritin and interleukin-6 decreased significantly after treatment with Remdesivir, with p-value <0.01, as compared to values at the time of admission, without any significant side effects.

Conclusion: Early Remdesvir administration demonstrated significant efficacy in moderate to severe COVID-19 patients hospitalized in ICU, reducing time to clinical improvement and mortality rate. It has been shown in previous studies to be of significant use in managing moderate to severe COVID-19 and reducing the mortality rate. As cases of COVID-19 are still rampant in our country it may be used in future practice.

 

Author (s) Details

Jyoti Prakash
Department of General Medicine, AIIMS Patna, India.

 

B Das
Department of Medicine, Sri Krishna Medical College and Hospital (SKMCH), Muzaffarpur, Bihar, India.

 

 

Please see the book here:- https://doi.org/10.9734/bpi/mmrnp/v8/2421

 

A Review of FTIR Spectroscopy Used in COVID-19 Analysis | Chapter 5 | Medical Science: Recent Advances and Applications Vol. 5

The worldwide risk that the coronavirus pandemic presents to millions of people is widespread. The cause of this pandemic, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was initially determined in Wuhan, the capital of Hubei province, and it has since spread to other parts of the world. Early coronavirus identification improves the possibility of infection treatment and inhibits the virus's transmission. The early detection and diagnosis of COVID-19 has made extensive use of spectroscopic methods, including infrared spectroscopy. Fourier transform infrared spectroscopy can identify changes in the biochemical events occurring in cells before and after a viral infection. It detects molecular vibrations caused by shifts in the electric dipole moment in chemical bonds, which are brought about by light absorption in the electromagnetic spectrum's 400–5000 cm-1 infrared region. However, because each spectral detection category uses a different detection technique, their sensitivity and specificity change. This review focuses on recent uses of FTIR spectroscopy in the detection of COVID-19.

 

Author (s) Details

Andrei A. Bunaciu
S.C. AAB IR Research S.R.L., Bragadiru – Ilfov District, 077025, Romania.

 

Hassan Y. Aboul-Enein
Pharmaceutical and Medicinal Chemistry Department, Pharmaceutical and Drug Industries Research Division, National Research Center, Cairo 12622, Egypt.

 

 

Please see the book here:- https://doi.org/10.9734/bpi/msraa/v5/5474

Semi-Quantitative Analysis of PAX6 Gene in Ocular and Non-Ocular Tissues | Chapter 6 | New Visions in Medicine and Medical Science Vol. 3

 PAX6 is a candidate gene for the proper development of human eye. The gene encompasses on the chromosome number 11p13. The expression of PAX6 gene occurs not only in eye but in many human organs like brain and pancreas. The present study focuses on the transcript analysis in ocular tissues and non-ocular tissues.

 

The total mRNA extracted from ocular tissues including lens epithelium, retina, iris and the non-ocular tissues like Trabecular Meshwork Cells, HeLa Cells, besides Aniridic patient as well as the normal person blood samples. The isolated RNAs were subjected to convert into cDNA by RT-PCR along with the house keeping genes (beta actin) used as a positive control in all the reactions.

 

The post PCR analysis revealed 540bp amplicon in all the isolated ocular and non-ocular tissues thus proved that the reaction setup was correct. In the ocular tissues showed approximately 600bp in retina, 855bp in iris and 1055bp in lens epithelium (PAX6 and PAX6 5a isoforms). The expression analysis of PAX6 gene in Non-ocular tissues HeLa cells showed 1.2 kb amplification however the mutant revealed showed unusual banding pattern of 190bp and 200bp. Sequencing of the unusual bands showed the irrelevant hypothetical LOC441763 mRNA. RT-PCR amplification confirms the presence of various PAX6 mRNA expressions in ocular and non-ocular tissues.

 

Lens epithelium, retina and iris tissues have significant PAX6 expression while the retina contains an overwhelming majority of mRNAs encoding crystalline and other genes but still express PAX6 isoforms at high levels. HeLa is a fibroblast based cell line and it would not expect a fibroblast to express a lot of PAX6 However PAX6 gene expression might be high in these cells for some reason. The PAX6 gene partial expression was found in mutant WBC and not in wild type WBC cells, the real fact for this reason is unknown. Hence the exploration of the PAX6 gene should be further analyzed in future.

Author(s) Details:

G. Neethirajan,
Department of Genetics, Aravind Medical Research Foundation, India and PG Research Centre in Biotechnology, MGR College, Hosur, TN, India.

Krishnadas S. R.,
Department of Glaucoma, Aravind Eye Hospital, India.

Vijayalakshmi P.,
Department of Pediatric Ophthalmology and Strabismus, Aravind Eye Hospital, India.

Shashikanth Shetty,
Department of Pediatric Ophthalmology and Strabismus, Aravind Eye Hospital, India.

Sundaresan P.,
Department of Genetics, Aravind Medical Research Foundation, India.

Please see the link here: https://stm.bookpi.org/NVMMS-V3/article/view/13954

Expression of Prostate-Specific Genes (STAMP1/STEAP2) in the Early Stages of Prostate Cancer| Chapter 9 | Novel Aspects on Pharmaceutical Research Vol. 7

 This branch focused on prostate-specific genes articulated in the early stages of prostate malignancy. Prostate cancer is the second most frequent cancer disease made in fathers and the fifth leading cause of decease worldwide. Prostate malignancy may be asymptomatic at the early stage and frequently has an indolent course that may demand only active following.  Prostate cancer may be asymptomatic at the beginning and often has an lazy course, and may require slightest or even no treatment. However, ultimate frequent complaint is difficulty accompanying urination, increased commonness, and nocturia, all symptoms that concede possibility also arise from prostatic hypertrophy. The belongings of androgens are mediated apiece androgen receptor (AR). Therefore, studies focus on identification of AR controlled genes that are also well expressed in the prostate. STAMP family genes STAMP1/STEAP2 [1] and STAMP2/STEAP4 [2] are only signified in androgen receptor positive cells, the function of AR in STAMP family deoxyribonucleic acid expression is an important question. Apart from the prostate, STAMP1 is in the direction of the heart, intellect, pancreas, ovary, skeleton, influence, mammary gland, testis, uterus, kind, lung, trachea, and liver. RT-PCR was used to determine an apoptosis panel including pro-apoptotic and/or apoptotic chemicals in LNCaP containers expressing STAMP genes per se.  The increase activities of STAMP1 give the impression linked to the ERK (extracellular signal-regulated kinase) road. Although the STAMP1/STEAP2 gene supporter region lacks androgen receptor response piece (ARRE), the second found STAMP2/STEAP4 deoxyribonucleic acid does have ARRE, as demonstrated by ChIP.

Author(s) Details:

Ceren Gönen,
Department of Pharmacology, Ege University, Faculty of Pharmacy, Bornova, Izmir, Turkey.

Please see the link here: https://stm.bookpi.org/NAPR-V7/article/view/11439

Studies on Viral Respiratory Infections with Bocavirus in Romania | Chapter 8 | Research Advances in Microbiology and Biotechnology Vol. 5

 Tobias Allander and others. (Karolinska University Hospital, Stockholm, Sweden) first identified the human Bocavirus (BoV) in nasopharyngeal aspirates captured from children with harsh respiratory sicknesses in 2001. Using a random PCR cloning sequencing approach, it was raise that 17 out of 540 samples (3.1%) held a novel DNA virus. The name Bocavirus is derived from member of genus bos and canine, refer to the known hosts for other appendages of this type: the bovine parvovirus, which infects oxen, and the minute virus of dogs, which infects dogs. The virus's ghost in serum, seat, and respiratory secretions plans that it may cause systemic sickness.Bocavirus is a single-marooned DNA virus that is a appendage of the Parvoviridae family. The samples secondhand in our investigation were nasopharyngeal exudates taken from 309 victims, aged 19 days to 96 age, who were hospitalized middle from two points September 2011 and September 2012 with a disease of SARI (severe acute respiring infections).Multiplex RT-PCR (reverse copy-polymerase chain reaction) with the Seeplex 15 One-Step RV ACE Detection Kit was the demonstrative technique applyied to find human BoV. Ten of the 309 samples tested for human bocavirus were found expected positive. Children (7 months to 3.5 age old) accompanying bronchiolitis and pneumonia without gastrointestinal symptoms given all 10 samples. The data reasoning revealed the following findings: HumanBoV is still present in Romania, meeting accompanying young children. The percent of 3.3% is comparable to      that found in Sweden but inferior in Jordan (9%), China (7%) and Taiwan (5, 6%).Patients admitted to the hospital and pronounced with BoV had only respiring symptoms (LRTI, lower respiratory lot infections) and no digestive syndromes.Cases of BoV positivity were not linked to added viruses.

Author(s) Details:

Cristina Tecu,
NIRDMI Cantacuzino, Romania.

Maria Elena Mihai,
NIRDMI Cantacuzino, Romania.

Viorel Alexandrescu,
NIRDMI Cantacuzino, Romania.

Alina Ivanciuc,
NIRDMI Cantacuzino, Romania.

Gheorghe Necula,
NIRDMI Cantacuzino, Romania.

Emilia Lupulescu,
NIRDMI Cantacuzino, Romania.

Odette Popovici,
National Center for Communicable Disease Surveillance and Control, Romania.

Please see the link here: https://stm.bookpi.org/RAMB-V5/article/view/10486