scholarly journals Optimization and validation of RT-LAMP assay for diagnosis of SARS-CoV2 including the globally dominant Delta variant

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Vijay Lakshmi Jamwal ◽  
Natish Kumar ◽  
Rahul Bhat ◽  
Piyush Singh Jamwal ◽  
Kaurab Singh ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Binding Sites ◽  
Lamp Assay ◽  
World Health ◽  
Molecular Testing ◽  
Double Blind Study ◽  
Double Blind ◽  
Common Technique ◽  
Primer Sets ◽  
Health Organization

Abstract Background Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the causative agent of COVID-19 pandemic, has infected more than 179 million people worldwide. Testing of infected individuals is crucial for identification and isolation, thereby preventing further spread of the disease. Presently, Taqman™ Reverse Transcription Real Time PCR is considered gold standard, and is the most common technique used for molecular testing of COVID-19, though it requires sophisticated equipments, expertise and is also relatively expensive. Objective Development and optimization of an alternate molecular testing method for the diagnosis of COVID-19, through a two step Reverse Transcription Loop-mediated isothermal AMPlification (RT-LAMP). Results Primers for LAMP were carefully designed for discrimination from other closely related human pathogenic coronaviruses. Care was also taken that primer binding sites are present in conserved regions of SARS-CoV2. Our analysis shows that the primer binding sites are well conserved in all the variants of concern (VOC) and variants of interest (VOI), notified by World Health Organization (WHO). These lineages include B.1.1.7, B.1.351, P.1, B.1.617.2, B.1.427/B.1.429, P.2, B.1.525, P.3, B.1.526 and B.1.617.1. Various DNA polymerases with strand displacement activity were evaluated and conditions were optimized for LAMP amplification and visualization. Different LAMP primer sets were also evaluated using synthetic templates as well as patient samples. Conclusion In a double blind study, the RT-LAMP assay was validated on more than 150 patient samples at two different sites. The RT-LAMP assay appeared to be 89.2% accurate when compared to the Taqman™ rt-RT-PCR assay.

Giant Primary Neuroendocrine Neoplasms of the Liver: Report of 2 Cases With Molecular Characterization

2019 ◽  
Vol 27 (8) ◽  
pp. 893-899
Author(s):  
Laura G. Pastrián ◽  
Ignacio Ruz-Caracuel ◽  
Raul S. Gonzalez
Keyword(s):  
World Health Organization ◽  
Neuroendocrine Tumor ◽  
Large Cell ◽  
The Other ◽  
World Health ◽  
Neuroendocrine Neoplasms ◽  
Molecular Testing ◽  
Well Differentiated ◽  
Grade 3 ◽  
Health Organization

Primary neuroendocrine neoplasms of the liver have occasionally been reported in the liver, though many reports do not convincingly exclude metastases. In this article, we report 2 “giant” hepatic neuroendocrine lesions without evidence of a primary elsewhere after clinical workup. One occurred in a 21-year-old male; the lesion was a large cell neuroendocrine carcinoma measuring 24 cm. The patient died of disease in 10 months. The other occurred in a 25-year-old patient, was 18 cm wide, and was diagnosed as a well-differentiated neuroendocrine tumor, World Health Organization grade 3. The patient died of disease after 30 months. Molecular testing demonstrated only the presence of TP53 mutations in common. These cases expand our knowledge of seemingly primary neuroendocrine neoplasms of the liver, in particular, giant cases measuring more than 8 cm. Guidelines for clinical workup and therapy for these lesions remain unclear, but future thorough workup of such cases is necessary for specific characterization.

scholarly journals Rifabutin-Containing Triple Therapy (RHB-105) for Eradication of Helicobacter pylori: Randomized ERADICATE Hp Trial

Antibiotics ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 685 ◽  
Author(s):  
Ira N. Kalfus ◽  
David Y. Graham ◽  
Dennis S. Riff ◽  
Raymond M. Panas
Keyword(s):  
Helicobacter Pylori ◽  
Eradication Rate ◽  
Study Drug ◽  
Standard Of Care ◽  
Empiric Therapy ◽  
World Health ◽  
Double Blind Study ◽  
Double Blind ◽  
Treatment Naïve ◽  
H Pylori

Due to increasing resistance to commonly used antibiotics, the World Health Organization and Food and Drug Administration have advocated the development of new therapeutic regimens for Helicobacter pylori (H. pylori). This phase three, double-blind study (ERADICATE Hp) randomized (2:1) treatment-naïve adults with H. pylori infection and dyspepsia to RHB-105 (an all-in-one combination of omeprazole 40 mg, amoxicillin 1000 mg, and rifabutin 50 mg) or an identically-appearing placebo, both administered every 8 h for 14 days. The H. pylori eradication rate with RHB-105, using a modified intent-to-treat (mITT) population of subjects who received ≥1 dose of study drug and had test-of-eradication performed 28–35 days post-completion of therapy, was compared (one-sample Z-test) to a literature-derived comparator rate of 70% and success rate with physician-selected standard-of-care given to placebo failures. The mITT H. pylori eradication rate (95% CI) with RHB-105 of 89.4% (82.0–96.8%) was greater than both the literature-derived comparator rate (P < 0.001) and the standard-of-care rate of 63.0% (44.8–81.1%) (P = 0.006). Adverse events with an incidence ≥5% for RHB-105 were diarrhea (12.7%), headache (11.9%), chromaturia (9.3%), abdominal tenderness (6.8%), and dizziness (5.1%). No leukopenia was noted. RHB-105 (Talicia®) proved to be a safe and effective empiric therapy for H. pylori eradication.

scholarly journals Advances in Diagnostic Methods for Zika Virus Infection

2018 ◽  
Vol 12 (4) ◽  
Author(s):  
Carlos A. Herrada ◽  
Md. Alamgir Kabir ◽  
Rommel Altamirano ◽  
Waseem Asghar
Keyword(s):  
Reverse Transcription ◽  
Zika Virus ◽  
Vaccine Development ◽  
Genome Structure ◽  
Diagnostic Methods ◽  
World Health ◽  
Health Concern ◽  
Localized Surface Plasmon ◽  
Enzyme Linked Immunosorbent Assay ◽  
Health Organization

The Zika virus (ZIKV) is one of the most infamous mosquito-borne flavivirus on recent memory due to its potential association with high mortality rates in fetuses, microcephaly and neurological impairments in neonates, and autoimmune disorders. The severity of the disease, as well as its fast spread over several continents, has urged the World Health Organization (WHO) to declare ZIKV a global health concern. In consequence, over the past couple of years, there has been a significant effort for the development of ZIKV diagnostic methods, vaccine development, and prevention strategies. This review focuses on the most recent aspects of ZIKV research which includes the outbreaks, genome structure, multiplication and propagation of the virus, and more importantly, the development of serological and molecular detection tools such as Zika IgM antibody capture enzyme-linked immunosorbent assay (Zika MAC-ELISA), plaque reduction neutralization test (PRNT), reverse transcription quantitative real-time polymerase chain reaction (qRT-PCR), reverse transcription-loop mediated isothermal amplification (RT-LAMP), localized surface plasmon resonance (LSPR) biosensors, nucleic acid sequence-based amplification (NASBA), and recombinase polymerase amplification (RPA). Additionally, we discuss the limitations of currently available diagnostic methods, the potential of newly developed sensing technologies, and also provide insight into future areas of research.

scholarly journals Ultrasound-Guided Pectoral Nerve Block With Tr Ansverse Thoracic Muscle Plane Block Enhanced World Health Organization on Quality of Life Brief Scale Scores 6 Months After M Odified Radical Mastectomy : A Randomized, Placebo-controlled and Double-Blind Clinical Trial

2021 ◽  
Author(s):  
Tianxiao Liu ◽  
Lina Chen ◽  
Min Shi ◽  
Weixin Dai ◽  
Jing Chen ◽  
...  
Keyword(s):  
Quality Of Life ◽  
Chronic Pain ◽  
World Health Organization ◽  
Radical Mastectomy ◽  
World Health ◽  
Ultrasound Guided ◽  
Double Blind ◽  
Thoracic Muscle ◽  
Health Organization

Abstract Background Modified radical mastectomy (MRM) has a large incision range and can cause strong intraoperative stress, a high incidence of postoperative acute pain, and chronic pain. There are few studies on the objective evaluation of the perioperative stress response by some stress-related serological indicators and on the long-term follow-up evaluation of postoperative quality of life and the incidence of chronic pain. This study aimed to evaluate the efficacy of ultrasound guided type Ⅰ Pecs block (Pecs Ⅰ), type Ⅱ Pecs block (Pecs Ⅱ) and transverse thoracic muscle plane block (TTP) on reducing CRP, IL-6, and WBC values during surgery and on enhancing World Health Organization on Quality of Life Brief Scale (QOL) scores 6 months after MRM.Methods The randomized, placebo-controlled and double-blind study was conducted in 76 patients assigned to two groups that received either ultrasound guided Pec Ⅰ + Pec Ⅱ +TTP block with 40 ml of 0.25% ropivacaine (group PT) or saline (group C). The primary outcomes were the changes in CRP, IL-6 and WBC values on the first day before surgery and the first and third days after surgery, the changes in blood glucose levels before and after surgery, and the QOL scores evaluated 6 months after surgery.Results The median (IQR) CRP, IL-6, and WBC values were significantly reduced in group PT on the first day compared with those in group C (12 (10-13.25) mg/l, 10.45 (9.575-11.65) pg/ml, 9 (7.75-9.25)×109/ l, vs 24 (20.75-26.25) mg/l, 25.35 (19.3-29.675) pg/ml, 12 (11-13) ×109/l, respectively, p< 0.001). The median (IQR) QOL was significantly higher in group PT 6 months after surgery than in group C (53(51.75-55) vs 43 (41-51.5), p< 0.001). The incidence of chronic pain was significantly lower, n/all was 4/33 vs 0/34, p< 0.001. Conclusions Ultrasound guided Pecs I, II and TTP block, significantly reduced the CRP, IL-6 and WBC values on the first day; increased the QOL scores and the incidence of chronic pain 6 months after surgery; and promoted the rapid recovery of patients.Trial registration: chictr.org.cn ChiCTR2000033275. Registered on 26 May 2020

scholarly journals Updates on 2016 CNS tumor classification - Where do we stand?

2019 ◽  
Vol 1 (1) ◽  
pp. 31-34
Author(s):  
Manish Kolakshyapati ◽  
Tejashwi Shrestha ◽  
Anish Man Singh ◽  
Rupendra Bahadur Adhikari ◽  
Prabin Shrestha
Keyword(s):  
International Standards ◽  
World Health ◽  
Patient Treatment ◽  
Molecular Testing ◽  
Diffuse Glioma ◽  
Treatment Protocols ◽  
Cns Tumor ◽  
New Treatment ◽  
Health Organization ◽  
Experimental Trials

The updated 2016 World Health Organization (WHO) classification of central nervous system (CNS) tumors has incorporated the molecular markers together with histological parameters in defining and diagnosing various tumor entities. This has led to major changes and revision of the classification. A number of new entities are classified based on clinical, diagnostic, and prognostic relevance and those without such relevance have been removed. Major changes are incorporated in diffuse glioma, medulloblastoma, pediatric gliomas, and other embryonal tumors. These new requisites make molecular testing a basic requirement for CNS tumor diagnosis and treatment. New treatment protocols are designed and targeted based on the genetic alteration involved in the tumor entity. The update aims to achieve accurate diagnosis, determine precise prognosis and ensure better patient treatment all through facilitating better categorization of clinical and experimental trials. The multidisciplinary team of clinicians treating such tumor patients in a developing country like Nepal should try to adopt this updated version of the classification so as to improve and upgrade the neuro-oncological services to meet the international standards.

Comparison of the NeuMoDX, Diasorin Simplexa, Cepheid and Roche CDC SARS-CoV 2 EUA assays using nasopharyngeal/nasal swabs in universal transport media (UTM) and sputum and tracheal aspirates

2020 ◽  
Author(s):  
Robert Tibbetts ◽  
Kathy Callahan ◽  
Kareem Rofoo ◽  
Richard J. Zarbo ◽  
Linoj Samuel
Keyword(s):  
The United States ◽  
World Health ◽  
Molecular Testing ◽  
Additional Specimen ◽  
Significant Difference ◽  
Nasal Swabs ◽  
Henry Ford ◽  
United States Food ◽  
Health Organization ◽  
Tracheal Aspirates

AbstractIn March 2019 the outbreak of SARS-CoV 2 was officially defined as a pandemic by the World Health Organization and shortly after, the United States Food and Drug Administration (FDA) granted Emergency Use Authorization (EUA) to the Centers for Disease Control (CDC) for reverse transcription polymerase chain reaction (rtPCR) molecular testing for the detection of the SARS-CoV-2 virus from NP swabs. Since then, EUA with relaxed regulations were granted to numerous manufacturers and clinical microbiology laboratories to implement in-house testing assays with nasopharyngeal swabs (NP) and subsequently additional specimen types. Because of supply chain shortages leading to competition for reagents, sustaining any significant volume of testing soon became problematic. As a countermeasure, within several weeks the Henry Ford Microbiology Laboratory validated 4 different rtPCR assays and multiple specimen types using NeuMoDX, Diasorin Simplexa, Cepheid and Roche platforms. The purpose of this study was to analyze the analytic sensitivity of these rtPCR assays with NP/nasal swabs and sputum/tracheal aspirates. Qualitative analytic agreement between the 4 platforms for NP/nasal swabs ranged 95% - 100% overall with no statistically significant difference in threshold cT values. Similar results were obtained with the sputum/tracheal aspirates. These data demonstrate the high accuracy and reproducibility in detection of SARS-CoV 2 between the rtPCR assays performed on 4 different platforms with numerous specimen types.

scholarly journals Early Treatment of COVID-19 Pneumonia with Anakinra Guided by Urokinase Plasminogen Receptor

2021 ◽  
Author(s):  
Evangelos Giamarellos-Bourboulis ◽  
Evdoxia Kyriazopoulou ◽  
Garyfallia Poulakou ◽  
Haralampos Milionis ◽  
Simeon Metallidis ◽  
...  
Keyword(s):  
Early Recognition ◽  
Severe Disease ◽  
World Health ◽  
Clinical Progression ◽  
Urokinase Plasminogen Activator Receptor ◽  
Double Blind ◽  
Respiratory Dysfunction ◽  
Once Daily ◽  
Plasminogen Activator Receptor ◽  
Health Organization

Abstract Early recognition of risk and start of treatment may improve unfavorable outcome of COVID-19. In the SAVE-MORE double-blind randomized trial, 594 patients with pneumonia without respiratory dysfunction at risk as defined by plasma suPAR (soluble urokinase plasminogen activator receptor) ≥ 6 ng/ml were 1:2 randomized to subcutaneous placebo or 100 mg anakinra once daily for 10 days; 85.9% were co-administered dexamethasone. After 28 days, anakinra-treated patients were distributed to lower strata of the 11-point World Health Organization ordinal Clinical Progression Scale (WHO-CPS) (adjusted odds ratio-OR 0.36; 95%CI 0.26–0.50; P < 0.001); anakinra protected from severe disease or death (≥ 6 points of WHO-CPS) (OR: 0.46; P: 0.010). The median WHO-CPS decrease in the placebo and anakinra groups was 3 and 4 points (OR 0.40; P < 0.0001); the median decrease of SOFA score was 0 and 1 points (OR 0.63; P: 0.004). 28-day mortality decreased (hazard ratio: 0.45; P: 0.045) and hospital stay was shorter. (Sponsored by the Hellenic Institute for the Study of Sepsis ClinicalTrials.gov identifier, NCT04680949)

Molecular testing in endometrial carcinoma – joint recommendation of Czech Oncological Society, Oncogynecological Section of the Czech Gynecological and Obstetrical Society, Society of Radiation Oncology, Biology and Physics, and the Society of Czech Pathologists

2021 ◽  
Vol 86 (4) ◽  
pp. 264-272
Author(s):  
Pavel Dundr ◽  
◽  
David Cibula ◽  
Martin Doležel ◽  
Pavel Fabián ◽  
...  
Keyword(s):  
Endometrial Carcinoma ◽  
European Society ◽  
Radiation Oncology ◽  
Molecular Classification ◽  
World Health ◽  
Molecular Testing ◽  
Therapeutic Implications ◽  
Health Organization ◽  
Endometrial Carcinomas

Summary: Molecular classification of endometrial carcinoma is becoming an important part of the dia gnostic process with direct therapeutic implications. Recent international guidelines, including the joint recommendation of the European Society of Gynaecological Oncology, the European Society for Radiotherapy and Oncology and the European Society of Pathology include the molecular classification into standard diagnostic algorithms. Molecular testing of endometrial carcinomas is also recommended in the latest (5th edition) of the World Health Organization classification of female genital tumors. Due to the need to implement these recommendations in practice, representatives of four professional societies of the Czech Medical Association of J. E. Purkyně (the Czech Oncological Society, the Oncogynecological Section of the Czech Gynecological and Obstetrical Society, the Society of Radiation Oncology, Biology and Physics, and the Society of Czech Pathologists) organized a meeting focused on this topic. Recommendation for molecular testing of endometrial carcinoma in routine dia gnostic practice in the Czech Republic.

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