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Isolation and Characterization of SARS-CoV-2 in Kenya

American Journal of Molecular Biology 2024年第2期14卷 66-83页

作者： Albina Makio Robinson Mugasiali Irekwa Matthew Mutinda Munyao caroline wangui njoroge Peter Kipkemboi Rotich Tonny Teya Nyandwaro Joanne Jepkemei Yego Anne Wanjiru Mwangi James Hungo Kimotho Ronald Tanui Vincent Rutto Samson Muuo Nzou Center for Traditional Medicine and Drug Research Kenya Medical Research Institute Nairobi Kenya Center for Microbiology Research Kenya Medical Research Institute Nairobi Kenya Production Department Kenya Medical Research Institute Nairobi Kenya Department of Biochemistry and Microbiology Rhodes University Grahamstown South Africa Center for Virus Research Kenya Medical Research Institute Nairobi Kenya

The discovery of Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) in Wuhan, Hubei province, China, in December 2019 raised global health warnings. Quickly, in 2020, the virus crossed borders and infected individuals across the world, evolving into the COVID-19 pandemic. Notably, early signs of the virus’s existence were observed in various countries before the initial outbreak in Wuhan. As of 12th of April, the respiratory disease had infected over 762 million people worldwide, with over 6.8 million deaths recorded. This has led scientists to focus their efforts on understanding the virus to develop effective means to diagnose, treat, prevent, and control this pandemic. One of the areas of focus is the isolation of this virus, which plays a crucial role in understanding the viral dynamics in the laboratory. In this study, we report the isolation and detection of locally circulating SARS-CoV-2 in Kenya. The isolates were cultured on Vero Cercopithecus cell line (CCL-81) cells, RNA extraction was conducted from the supernatants, and reverse transcriptase-polymerase chain reaction (RT-PCR). Genome sequencing was done to profile the strains phylogenetically and identify novel and previously reported mutations. Vero CCL-81 cells were able to support the growth of SARS-CoV-2 in vitro, and mutations were detected from the two isolates sequenced (001 and 002). Genome sequencing revealed the circulation of two isolates that share a close relationship with the Benin isolate with the D614G common mutation identified along the S protein. These virus isolates will be expanded and made available to the Kenya Ministry of Health and other research institutions to advance SARS-CoV-2 research in Kenya and the region.

关键词： SARS-CoV-2 COVID-19 Whole Genome Sequencing Phylogenetic Analysis Nucleotide Substitutions Amino Acid Changes

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Isolation and Characterization of SARS-CoV-2 in Kenya

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American Journal of Molecular Biology（美国分子生物学期刊） 2024年第2期 66页

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Expression and Evaluation of Wb-SXP-1 and Wb-123 Recombinant Antigens as Potential Diagnostic Biomarkers for Lymphatic Filariasis

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American Journal of Molecular Biology 2023年第2期13卷 95-112页

作者： Sharlene Kerubo Mageto Rebecca Wanjiku Waihenya Anne Wanjiru Mwangi Peter Kipkemboi Rotich Matthew Mutinda Munyao Tonny Teya Robinson Mugasiali Irekwa Joanne Jepkemei Yego caroline wangui njoroge Grace Ng’endo Kanyita Nicole Sian Tanchu Dawala Koromtili Oumar Primrose Muthoni Ndungu Samson Muuo Nzou Department of Molecular Biology and Biotechnology Pan-African University Institute of Basic Sciences Technology and Innovation Nairobi Kenya Zoology Department Jomo Kenyatta University for Agriculture and Technology Nairobi Kenya Nagasaki University Institute of Tropical Medicine Kenya Medical Research Institute (NUITM-KEMRI) Nairobi Kenya Production Department Kenya Medical Research Institute Nairobi Kenya Centre for Microbiology Research Kenya Medical Research Institute Nairobi Kenya Department of Biochemistry Microbiology and Biotechnology Kenyatta University Nairobi Kenya Department of Pharmacology Clinical Pharmacy and Pharmacy Practice University of Nairobi Nairobi Kenya

Lymphatic filariasis (LF) remains a public health concern as it can cause permanent morbidity and disability to those infected. While the global elimination of LF in these endemic areas is ongoing through mass drug administration, there is the need to develop diagnostic tools that would be utilized to track the progress of total global eradication as well as perform surveillance for the recurrence of lymphatic filariasis transmission. Currently, approved LF diagnosis tools are faced with lack of specificity, low sensitivity, and periodicity dependence. Recombinant filarial antigen-based assays can address these drawbacks and offer practical instruments for LF diagnosis and surveillance. This present study, evaluated rWb-SXP-1 and rWb-123 antigens as potential diagnostic biomarker tools for Wuchereria banchrofti in human sera using microspheres-based multiplex serological assay. Based on statistical analysis using XLSTAT 2019 (Addinsoft) on data generated from multiplex technology assay, generated ROC curves for both rWb-SXP-1 and rWb-123 demonstrated 87.1% sensitivity to Wuchereria banchrofti human sera with rWb-SXP-1 antigens having the highest specificity of 96%. Indication that rWb-SXP-1 and rWb-123 antigens are capable of detecting immunoglobulin G4 (IgG4) antibodies in human sera synthesized specifically against W. banchrofti infections. Therefore, rWb-SXP-1 and rWb-123 antigens can be utilized to detect W. banchrofti infections by antibody profiling with excellent diagnostic sensitivity and specificity using microsphere-based multiplex serological tests. This method can be particularly practical for screening a large number of sera samples and/or for quick, extensive field-testing due to the high-throughput and quick formats applied.

关键词： Lymphatic Filariasis Recombinant Antigens Diagnosis

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Investigation of Plasmodium falciparum Resistance Biomarkers among Primary School Children in Western Kenya

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American Journal of Molecular Biology 2022年第2期12卷 43-53页

作者： Otilmoi Poul Stephen Tonny Teya Nyandwaro Robinson Mugasiali Irekwa Rebecca Wanjiku Waihenya Matthew Mutinda Munyao Peter Kipkemboi Rotich caroline wangui njoroge Anne Wanjiru Mwangi Joanne Jepkemei Yego Nicole Sian Tanchu Dawala Koromtili Oumar Grace Ngendo Kanyita Primrose Muthoni Ndungu Samson Muuo Nzou Department of Molecular Biology and Biotechnology Pan-African University Institute of Basic Sciences Technology and Innovation Nairobi Kenya Innovation Technology Transfer Department Kenya Medical Research Institute (ITTD-KEMRI) Nairobi Kenya Zoology Department School of Biological Sciences College of Pure and Applied Sciences Jomo Kenyatta University of Agriculture and Technology Nairobi Kenya Production Department KEMRI Nairobi Kenya Centre for Microbiology Research Kenya Medical Research Institute Nairobi Kenya Nagasaki University Institute of Tropical Medicine Kenya Medical Research Institute (NUITM-KEMRI) Nairobi Kenya Department of Biochemistry Microbiology and Biotechnology Kenyatta University Nairobi Kenya Department of Pharmacology and Pharmacognosy University of Nairobi Nairobi Kenya

Background: A marked decrease in malaria-related deaths worldwide has been attributed to the administration of effective antimalarials against Plasmodium falciparum. However, the continuous spread of P. falciparum resistance to anti-malarial drugs is raising a serious problem in controlling Malaria to the vulnerable children’s immune system. In recent studies, Plasmodium falciparum Kelch 13 propeller gene (Pfk13) has been reported to develop resistance to artemisinin in South Asia. In this study, we checked Plasmodium falciparum chloroquine resistance transporter gene (Pfcrt) involved in chloroquine (CQ) resistance. Method: In this study, archived 280 samples were collected from Alupe primary school children in Busia, Western Kenya from May, 2016 to November, 2016. Genomic DNA was extracted using the MightyPrep reagent. The samples were investigated for P. falciparum positivity out of which 67 of them tested positive giving a prevalence rate of 24%. The sixty-seven were subjected to PCR amplification for the molecular marker resistance to Pfcrt. After PCR amplification, the amplicons were purified and sequenced using Sanger Sequencing. The sequence data were analyzed using BioEdit software to identify point mutations. Results: 14 samples sequences were analyzed on Bioedit software giving the following amino acid changes F76C, Y66H, L70A, Y58C, T59V, V65I, P67L, T81L, Y60S, Y66S, P67T and I71F). New mutations have been reported at position 76 leading to an amino acid change, one of Pfcrt gold standard biomarkers. However, amino acid changes Y66H, L70A, Y58C, T59V, V65I, P67L, T81L, Y60S, Y66S, P67T and I71F are newly reported giving an increase in Pfcrt prevalence of concern from zero to 5.0%. A phylogenetic evolutionary relationship was constructed as shown below. Generally, the results showed a continuous resistance of *** to Pfcrt which calls for robust continuous monitoring and surveillance. Conclusion: Due to the increase of the resistant Pfcrt gene pre

关键词： P. falciparum Chloroquine Drug Resistance Malaria P. falciparum Chloroquine Resistance Transporter Gene Mutations

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Use of Polyclonal Antibody for the Diagnosis of Human African Trypanosomiasis

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American Journal of Molecular Biology 2023年第2期13卷 127-139页

作者： Dawala Koromtili Oumar Matthew Mutinda Munyao Anne Wanjiru Mwangi Rebecca Wanjiku Waihenya Peter Kipkemboi Rotich Robinson Mugasiali Irekwa Tonny Teya Nyandwaro caroline wangui njoroge Joanne Jepkemei Yego Primrose Muthoni Ndungu Sharlene Kerubo Mageto Damaris Mutethya Kilei Otilmoi Poul Stephen Nicole Sian Tanchu Grace Ngendo Kanyita Shingo Inoue Lucy Gitau Samson Muuo Nzou Department of Molecular Biology and Biotechnology Pan-African University Institute of Basic Sciences Technology and Innovation Nairobi Kenya Centre for Microbiology Research Kenya Medical Research Institute Nairobi Kenya Nagasaki University Institute of Tropical Medicine-Kenya Medical Research Institute Project Nairobi Kenya Production Department Kenya Medical Research Institute Nairobi Kenya Zoology Department Jomo Kenyatta University for Agriculture and Technology Nairobi Kenya Department of Pharmacology and Pharmacognosy University of Nairobi Nairobi Kenya Department of Microbiology Jomo Kenyatta University for Agriculture and Technology Nairobi Kenya Department of Biochemistry Microbiology and Biotechnology Kenyatta University Nairobi Kenya Center for Biotechnology Research and Development Kenya Medical Research Institute Nairobi Kenya

Human African trypanosomiasis (HAT), commonly known as sleeping sickness is one of the neglected tropical diseases (NTDs), which is fatal if left untreated. Its diagnosis is a challenge since the signs and symptoms of the primary phase are not specific, the existing diagnostic methods have low sensitivity and specificity, and the available drugs have some toxicity. New, robust, and cost-effective techniques are needed for the early identification of parasites. This study aimed to assess the sensitivity and specificity of two different types of polyclonal antibodies against T. b. gambiense using antigen detection ELISA. Polyclonal antibodies against the expressed proteins Tbg I2 and Tbg I17 were produced using New Zealand white rabbits. The antibody titer measured was greater than 32 g/L after the 3rd immunization for the expressed protein Tbg I2. For the expressed protein Tbg I17, the antibody titer measured was greater than 32 g/L after the 4th immunization. The sensitivity and specificity of the Tbg I2 polyclonal antibody confirmed with Polymerase Chain Reaction (PCR) as gold standard were respectively 89.5% and 80.6%, while for the Tbg I17 polyclonal antibody, the sensitivity and specificity were respectively 92.1% and 88.9%. The area under the curve for the Tbg I2 polyclonal antibody was 0.90 ± 0.032, while for the Tbg I17 polyclonal antibody, the area under the curve was 0.92 ± 0.0. The Tbg I17 polyclonal antibody produced in New Zealand white rabbits has good sensitivity and good specificity;it can be successfully used in the diagnosis of HAT.

关键词： Human African Trypanosomiasis Polyclonal Antibody Tbg I2 Expressed Protein Tbg I17 Expressed Protein Sensitivity Specificity

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Expression and Evaluation of Wb-SXP-1 and Wb-123 Recombinant Antigens as Potential Diagnostic Biomarkers for Lymphatic Filariasis

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American Journal of Molecular Biology（美国分子生物学期刊） 2023年第2期

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Use of Polyclonal Antibody for the Diagnosis of Human African Trypanosomiasis

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American Journal of Molecular Biology（美国分子生物学期刊） 2023年第2期

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物流管理对肯尼亚肯尼亚医疗用品管理局（KEMSA）的影响

物流管理对肯尼亚肯尼亚医疗用品管理局（KEMSA）的影响

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作者： NELLY wangui njoroge 北京交通大学

学位级别：硕士

本研究的旨在分析物流管理对肯尼亚肯尼亚医疗用品管理局（KEMSA）绩效的影响。该研究旨在实现以下具体目标:回顾和研究运输管理和订单管理如何影响肯尼亚肯尼亚医疗用品管理局的绩效,回顾和分析肯尼亚医疗用品管理局的仓储管理,调查库... 详细信息

本研究的旨在分析物流管理对肯尼亚肯尼亚医疗用品管理局（KEMSA）绩效的影响。该研究旨在实现以下具体目标:回顾和研究运输管理和订单管理如何影响肯尼亚肯尼亚医疗用品管理局的绩效,回顾和分析肯尼亚医疗用品管理局的仓储管理,调查库存管理如何影响肯尼亚医疗用品管理局的业绩第二章文献将从工作论文,期刊,书籍,报告,期刊和互联网资源等角度,回顾和批判性讨论以往的研究理论,同时指出研究差距。第三章将解释采用是研究方法。本研究旨在确定物流管理对肯尼亚肯尼亚医疗用品管理局（KEMSA）业绩的影响,涉及数据收集、测量和分析,本章讨论用于数据收集,处理和分析的程序和技术。第四章将呈现研究结果。研究运输管理、订单管理、仓储管理和库存管理四个方面如何影响肯尼亚肯尼亚医疗用品管理局（KEMSA）的绩效。本章重点关注数据分析,回答提出的研究问题。第五章的目标是分析讨论第四章提出的定性分析结果,同时得出结论和提出建议。

关键词：物流管理肯尼亚医疗用品管理局组织绩效运输管理订单管理仓库管理库存管

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Perioperative Awareness after Anesthesia: A Case Report

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Open Journal of Anesthesiology 2021年第5期11卷 149-155页

作者： caroline wangui Njue Ngugi M. Kinyungu St George’s University Grenada ITAV Pain Management Donald and Barbara Zucker School of Medicine at Hofstra/Northwell New York USA

Background: Accidental awareness during general anesthesia (AAGA) is an uncommon condition with possible long-term sequela. Here, we identify and describe a case of awareness in the perioperative setting and the resulting sequelae of symptoms consistent with post-traumatic stress disorder using the first-hand experience of a patient with awareness during anesthesia. Aim: This case report aims to identify and describe the symptomatic sequelae of perioperative awareness events and the importance of close patient follow-up and management. Case Presentation: A 34-year-old female developed uncontrolled tonsillar hemorrhage 2 days after successful adenotonsillectomy that necessitated bedside intubation. During regular clinic follow-up, the patient presented with symptoms suggestive of post-traumatic stress disorder (PTSD) secondary to recall events while under anesthesia. Conclusion: The contributory factors predisposing to perioperative awareness in the surgical environment need to be understood and avoided as much as possible. When awareness is suspected or known to have occurred the patient needs to be assisted through the post-op period and guided through the psychological sequelae that follow.

关键词： Accidental Awareness during General Anesthesia Post-Traumatic Stress Disorder

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Perioperative Awareness after Anesthesia: A Case Report

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Open Journal of Anesthesiology（麻醉学期刊） 2021年第5期

作者： caroline wangui Njue Ngugi M.Kinyungu

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