Article
Chemistry, Analytical
Ian T. Hull, Enos C. Kline, Gaurav K. Gulati, Jack Henry Kotnik, Nuttada Panpradist, Kamal G. Shah, Qin Wang, Lisa Frenkel, James Lai, Joanne Stekler, Barry R. Lutz
Summary: The study presents a novel quantitative detection method for HIV DNA or RNA using a combination of recombinase polymerase amplification and lateral flow detection technologies, allowing for rapid and accurate testing in low-resource healthcare settings, with the potential to improve the quality of HIV treatment.
ANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Analytical
Mengyun Zhou, Chuan Fan, Lirong Wang, Tailin Xu, Xueji Zhang
Summary: This study reports an integrated microdroplet array platform composed of an ultrasonic unit and minipillar array for enhanced RPA detection of SARS-CoV-2. The independent microvolume reactions and microstreaming driven by ultrasound greatly accelerate the amplification and increase the sensitivity, while reducing reagent consumption.
ANALYTICAL CHEMISTRY
(2022)
Article
Immunology
Bo Zhu, Juan Hu, Xuelian Li, Xiaomin Li, Lei Wang, Shihui Fan, Xin Jin, Kun Wang, Weiguo Zhao, Wenjun Zhu, Cheng Chen, Zilu Wang, Yingzhi Lu
Summary: Enterococcus faecalis is a serious issue in hospitals, necessitating the development of rapid and accurate detection methods. In this study, a visualized isothermal amplification method using RPA-LFS was developed for E. faecalis detection, showing fast, accurate, specific, and compatible results.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2022)
Article
Medicine, General & Internal
Md Anik Ashfaq Khan, Khaledul Faisal, Rajashree Chowdhury, Prakash Ghosh, Faria Hossain, Manfred Weidmann, Dinesh Mondal, Ahmed Abd El Wahed
Summary: This study developed a quantification method to assess the absolute quantification ability of isothermal recombinase polymerase amplification (RPA) assay for parasite genomes. The findings suggest the potential of Q-RPA assay in monitoring parasite load and developing rapid assays for other infectious diseases.
Article
Biotechnology & Applied Microbiology
Xiaohan Yang, Panpan Zhao, Yu Dong, Shiqi Chen, Hui Shen, Ge Jiang, Hai Zhu, Jingquan Dong, Song Gao
Summary: A new rapid detection method for V. vulnificus was developed using RPA technology and LFS strip, with good specificity and sensitivity, suitable for on-site detection.
Article
Immunology
Chao Ma, Shihui Fan, Yu Wang, Haitao Yang, Yi Qiao, Ge Jiang, Mingsheng Lyu, Jingquan Dong, Hui Shen, Song Gao
Summary: The study developed a real-time RPA assay for EHP detection, which showed rapid detection speed, good specificity, sensitivity comparable to PCR and LAMP methods, and significant improvement over the RPA assay analyzed by gel electrophoresis.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2021)
Article
Immunology
Fang Wang, Yan Wang, Xia Liu, Lei Wang, Kun Wang, Chenglai Xu, Guanhong Huang, Xuzhu Gao
Summary: In this study, a highly specific and sensitive diagnostic method for Streptococcus pneumoniae using the isothermal recombinase polymerase amplification (RPA) technique and gold-nanoparticle-based lateral flow strips (LFS) was developed. The RPA-LFS test showed high specificity and sensitivity for the identification of S. pneumoniae, especially in resource-limited areas.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2022)
Article
Agriculture, Dairy & Animal Science
Yanling Pang, Feng Cong, Xinheng Zhang, Hongxin Li, Yung-Fu Chang, Qingmei Xie, Wencheng Lin
Summary: A probe-based recombinase polymerase amplification assay was developed for rapid detection of Chlamydia psittaci. The test showed high sensitivity and accuracy in field applications, offering a promising approach for screening and monitoring outbreaks in domestic fowl populations.
Review
Biotechnology & Applied Microbiology
Xiaolei Mao, Minghui Xu, Shuyin Luo, Yi Yang, Jiaye Zhong, Jiawei Zhou, Huayan Fan, Xiaoping Li, Zhi Chen
Summary: In the field of pathogen detection, isothermal amplification technology is an important alternative to traditional PCR due to its speed, accuracy, and sensitivity. This paper explores the principles and current status of recombinase polymerase amplification (RPA) and recombinase-aid amplification (RAA), as well as the integration of the CRISPR-Cas system with these techniques. Furthermore, the paper reviews the combination of isothermal amplification and CRISPR-Cas technology and its practical applications in pathogen detection, highlighting the significant advancements it brings to research and clinical implementation.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)
Article
Chemistry, Multidisciplinary
Gaihua Cao, Jiangbo Dong, Xiaolong Chen, Peng Lu, Yifan Xiong, Lan Peng, Jiawei Li, Danqun Huo, Changjun Hou
Summary: In this study, a strategy (MPT-Cas12a/13a) was developed that combines CRISPR/Cas12a and Cas13a for simultaneous detection of CaMV35S and T-nos. This strategy utilizes multiplex PCR and transcription to achieve simultaneous detection with different signals in the same space. The MPT-Cas12a/13a showed excellent sensitivity, with detection limits as low as 11 copies of T-nos and 13 copies of CaMV35S, and demonstrated outstanding specificity and anti-interference ability in actual sample analysis. Therefore, it has the potential to be used for the detection of GM crops.
CHEMICAL COMMUNICATIONS
(2022)
Article
Plant Sciences
Kihye Shin, Soon-Hwa Kwon, Seong-Chan Lee, Young-Eel Moon
Summary: This study established a RPA-CRISPR/Cas12a-LFA system for rapid diagnosis of citrus scab, which is highly sensitive and does not cross-react with other fungal diseases.
Article
Virology
Lingjing Mao, Jiaxu Ying, Benjamin Selekon, Ella Gonofio, Xiaoxia Wang, Emmanuel Nakoune, Gary Wong, Nicolas Berthet
Summary: Monkeypox, caused by the monkeypox virus, is a zoonotic disease mainly found in West and Central Africa. Recent outbreaks in multiple countries worldwide have raised concerns about its global spread. The development of rapid diagnostic tests using recombinase-based isothermal amplification assays provides a promising solution for early detection and control of monkeypox outbreaks.
Article
Virology
Arianna Ceruti, Rea Maja Kobialka, Judah Ssekitoleko, Julius Boniface Okuni, Sandra Blome, Ahmed Abd El Wahed, Uwe Truyen
Summary: African swine fever virus (ASFV) is a deadly disease in pigs that spreads rapidly. A rapid detection assay using real-time recombinase polymerase amplification (RPA) for ASFV was developed and found to have equivalent sensitivity and specificity as real-time PCR. The combination of a simple extraction protocol allows for the use of the ASFV-RPA assay to improve control measures in outbreak situations.
Article
Chemistry, Analytical
Fangchi Shao, Joon Soo Park, Guojie Zhao, Kuangwen Hsieh, Tza-Huei Wang
Summary: Developing assays that combine CRISPR/Cas and isothermal nucleic acid amplification is a growing research area, with potential for point-of-care applications. Current research mainly focuses on appending different CRISPR/Cas effectors to isothermal nucleic acid amplification methods, but few studies have integrated both components into single-step assays. We conducted a systematic investigation and found that adding CRISPR/Cas12a enhances signal amplification but hinders recombinase polymerase amplification (RPA). We developed a PAM-free CRISPR/Cas12a-assisted RPA assay that outperforms other assays in design, signal, sensitivity, and speed. Our study provides new insights and can facilitate further advances in CRISPR/Cas-based assays.
ANALYTICAL CHEMISTRY
(2023)
Article
Biophysics
Ting Zheng, Xianming Li, Yanjun Si, Minjin Wang, Yuzhen Zhou, Yusheng Yang, Na Liang, Binwu Ying, Peng Wu
Summary: In this study, an accurate point-of-care testing (POCT) platform was proposed by specific recognition of target amplicons with peptide nucleic acid (PNA, assisted by T7 Exonuclease), which efficiently eliminated false positives and showed high sensitivity and specificity for detecting SARS-CoV-2.
BIOSENSORS & BIOELECTRONICS
(2023)
Article
Infectious Diseases
Prakash Ghosh, Rajashree Chowdhury, Mohammad Enayet Hossain, Faria Hossain, Mojnu Miah, Md Utba Rashid, James Baker, Mohammed Ziaur Rahman, Mustafizur Rahman, Xuejun Ma, Malcolm S. Duthie, Ahmed Abd El Wahed, Dinesh Mondal
Summary: The democratization of diagnostics is crucial for containing the transmission of COVID-19. This study evaluated two recombinase-based isothermal techniques, RT-RPA and RT-RAA, for detecting SARS-CoV-2. The results showed that both methods have high diagnostic accuracy and can be used for timely detection of the pathogen, understanding its epidemiology, case management, and transmission control.
INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES
(2022)
Article
Agriculture, Dairy & Animal Science
Sanaa M. Idris, Kamal H. Eltom, Julius B. Okuni, Lonzy Ojok, Wisal A. Elmagzoub, Ahmed Abd El Wahed, ElSagad Eltayeb, Ahmed A. Gameel
Summary: Paratuberculosis is a chronic disease caused by Mycobacterium avium subsp. paratuberculosis, affecting both ruminants and non-ruminants. It has been associated with Crohn's disease and is more prevalent in goats than in sheep. This disease is often neglected in poor communities where small ruminants play a vital role in people's livelihoods.
Review
Microbiology
Ellis Kobina Paintsil, Linda Aurelia Ofori, Sarah Adobea, Charity Wiafe Akenten, Richard Odame Phillips, Oumou Maiga-Ascofare, Maike Lamshoeft, Juergen May, Kwasi Obiri Danso, Ralf Krumkamp, Denise Dekker
Summary: This review summarizes the epidemiology and antibiotic resistance data of Campylobacter species isolated from humans and food-producing animals in West Africa. The study found a high prevalence of Campylobacter species in both humans and animals, with a higher rate of antibiotic resistance in human samples. Improved farm hygiene and surveillance systems are needed to reduce transmission.
Article
Multidisciplinary Sciences
Wisal A. Elmagzoub, Sanaa M. Idris, Maha Isameldin, Nassir Arabi, Abdelmonem Abdo, Mustafa Ibrahim, Md Anik Ashfaq Khan, Franziska Tanneberger, Sahar M. Bakhiet, Julius B. Okuni, Lonzy Ojok, Ahmed A. Gameel, Ahmed Abd El Wahed, Michael Bekaert, Mohamed E. Mukhtar, Ahmad Amanzada, Kamal H. Eltom, ElSagad Eltayeb
Summary: This study represents the first report of Mycobacterium avium subsp. paratuberculosis (MAP) in humans in Sudan, revealing a considerable proportion of the population may be infected or carriers of the bacteria. Increased awareness at the community level is urgently needed to reduce the risk of MAP transmission at the human-animal interface.
Article
Infectious Diseases
Noah Fongwen, Becca L. Handley, Diana L. Martin, Camila Beiras, Louise Dyson, Michael Frimpong, Oriol Mitja, Kingsley Asiedu, Michael Marks
Summary: New diagnostics are needed to aid the eradication of yaws. A working group has established target product profiles for developing new diagnostic tools based on identified use cases.
PLOS NEGLECTED TROPICAL DISEASES
(2022)
Article
Chemistry, Analytical
Md Anik Ashfaq Khan, Prakash Ghosh, Rajashree Chowdhury, Faria Hossain, Araf Mahmud, Abu S. G. Faruque, Tahmeed Ahmed, Ahmed Abd El Wahed, Dinesh Mondal
Summary: The need for fast detection of etiological agents beyond the target range of pathogens necessitates the use of rapid sequencing technologies. A PCR-free rapid nanopore barcoding assay was tested and found effective for detecting microbial species in acute diarrheal case specimens. Optimal parameters for detection accuracy were identified, and the method was found to have potential for widening the range of detectable pathogens in acute diarrheal samples.
ANALYTICAL CHEMISTRY
(2022)
Article
Multidisciplinary Sciences
Finja Rausch, Franziska Tanneberger, Ahmed Abd El Wahed, Uwe Truyen
Summary: The importance of air purifiers has increased, especially during the COVID-19 pandemic. This study aimed to establish a rapid molecular assay to differentiate intact and non-infectious virus particles, using a model virus. The study revealed a promising method to validate the efficacy of air purifiers by combining enzymatic pretreatment and molecular assays.
Article
Multidisciplinary Sciences
Shomik Maruf, Soumik Kha Sagar, Md. Masud Ur Rashid, Proggananda Nath, Md. Sahidul Islam, Prakash Ghosh, Md. Utba Rashid, Dinesh Mondal, Ahmed Abd El Wahed, Ariful Basher
Summary: This study discusses Para-KDL cases and their management, aiming to address gaps in existing methods of diagnosis and treatment.
Article
Infectious Diseases
Estelle Marion, Numfor Hycenth, Sundeep Chaitanya Vedithi, Marie Robbe-Saule, Valerie Donkeng, Line-Marlene Ganlonon, Affolabi Dissou, Solange Kakou Ngazoa, Marie-Jose Kabedi, Arsene Mabika Mabika, Richard Phillips, Michael Frimpong, Dorothy Yeboah-Manu, Vera Yatta Walker, Olaoluwa Akinwale, Maman Issaka, Gisela Bretzel, Kingsley Asiedu, Sara Eyangoh
Summary: Buruli ulcer is a neglected tropical disease mainly found in West and Central Africa. The mode of transmission is unknown and early diagnosis and treatment are crucial for reducing morbidity and preventing long-term disability. The gold standard for diagnosis is biological confirmation using PCR on cutaneous samples. In order to improve the diagnostic rate, 11 laboratories have created the network BU-LABNET to harmonize procedures and ensure quality control.
PLOS NEGLECTED TROPICAL DISEASES
(2022)
Article
Agriculture, Dairy & Animal Science
Sanaa M. M. Idris, Eva A. A. Ali, Wisal A. A. Elmagzoub, Julius B. B. Okuni, Mohamed E. E. Mukhtar, Lonzy Ojok, ElSagad Eltayeb, Ahmed Abd El Wahed, Kamal H. H. Eltom, Ahmed A. A. Gameel
Summary: This study reports the first occurrence of paratuberculosis in sheep in Sudan, highlighting the need for further investigations and control measures for this disease in the country.
Article
Microbiology
Chandrani Menike, Rajeewa Dassanayake, Renu Wickremasinghe, Maheeka Seneviwickrama, Indika De Alwis, Ahmed Abd El Wahed, Shalindra Ranasinghe
Summary: This study aimed to determine the risk of exposure to Leishmania infection among renal patients in Sri Lanka. The study found that the risk of Leishmania infection through blood transfusions was very low, suggesting that routine screening of kidney patients may not be necessary in Sri Lanka.
Article
Medicine, General & Internal
Oumar Ndiaye, Cheikh Tidiane Diagne, Ahmed Abd El Wahed, Fatou Dia, Moussa Dia, Adama Faye, Silvania Da Veiga Leal, Menilita dos Santos, Maria da Luz de Lima Mendonca, Carolina Cardoso da Silva Leite, Cheikh Saad Bouh Boye, Juliet E. Bryant, Philippe Despres, Ousmane Faye, Amadou Alpha Sall, Oumar Faye
Summary: This study describes an indirect ELISA for detecting specific IgG antibodies against ZIKV using the EDIII protein expressed in Drosophila S2 cells as an immunogen. The EDIII-ELISA showed a high sensitivity of 100.0% and specificity of 94.7% when compared to PRNTs, with 0.208 as the cut-off OD value. The recombinant EDIII can improve the sensitivity and specificity of IgG ELISA assay for ZIKV diagnostics.
Article
Virology
Arianna Ceruti, Ndongo Dia, Adeleye Solomon Bakarey, Judah Ssekitoleko, Soa Fy Andriamandimby, Padra Malwengo-Kasongo, Rasheeda H. A. Ahmed, Rea Maja Kobialka, Jean Michel Heraud, Moussa Moise Diagne, Marie Henriette Dior Dione, Idrissa Dieng, Martin Faye, Ousmane Faye, Jean Theophile Rafisandratantsoa, Jean-Pierre Ravalohery, Claudio Raharinandrasana, Tsiry H. Randriambolamanantsoa, Norosoa Razanajatovo, Iony Razanatovo, Joelinotahina H. Rabarison, Phillipe Dussart, Louis Kyei-Tuffuor, Abigail Agbanyo, Olubusuyi Moses Adewumi, Adeola Fowotade, Muideen Kolawole Raifu, Patient Okitale-Talunda, Gracia Kashitu-Mujinga, Christelle Mbelu-Kabongo, Steve Ahuka-Mundeke, Anguy Makaka-Mutondo, Enas M. Abdalla, Sanaa M. Idris, Wisal A. Elmagzoub, Rahma H. Ali, Eman O. M. Nour, Rasha S. M. Ebraheem, Huda H. H. Ahmed, Hamadelniel E. Abdalla, Musab Elnegoumi, Izdihar Mukhtar, Muatsim A. M. Adam, Nuha Y. I. Mohamed, Shahinaz A. Bedri, Hamdan Mustafa Hamdan, Magid Kisekka, Monica Mpumwiire, Sharley Melissa Aloyo, Joanita Nabwire Wandera, Andrew Agaba, Rogers Kamulegeya, Hosea Kiprotich, David Patrick Kateete, Paul Kadetz, Uwe Truyen, Kamal H. Eltom, Anavaj Sakuntabhai, Julius Boniface Okuni, Sheila Makiala-Mandanda, Vincent Lacoste, George Olusegun Ademowo, Michael Frimpong, Amadou Alpha Sall, Manfred Weidmann, Ahmed Abd El Wahed
Summary: In this study, a novel isothermal amplification method was validated for rapid detection of SARS-CoV-2 in seven sub-Saharan African countries. The RdRP-based amplification method showed the best assay accuracy. The study highlighted the challenges of implementing rapid molecular assays in field conditions, emphasizing the importance of standardized operation procedures, continuous training for staff, and enhanced quality control measures for successful deployment.
JOURNAL OF CLINICAL VIROLOGY
(2023)
Article
Infectious Diseases
Michael Frimpong, Venus Nana Boakyewaa Frimpong, Hycenth Numfor, Valerie Donkeng Donfack, Jennifer Seyram Amedior, Danielle Emefa Deegbe, Baaba Dadson, Anthony Ablordey, Sara M. Eyangoh, Richard Odame M. Phillips, Sundeep Chaitanya M. Vedithi
Summary: The gold standard for detection of Mycobacterium ulcerans is PCR, but the available PCR assays are not suitable for use in endemic communities due to a lack of mobility. We validated a dried reagent-based PCR assay for use with a portable thermocycler that could be used in communities with limited infrastructure. The assay showed high sensitivity and specificity, making it a suitable decentralized testing option for Buruli ulcer.
PLOS NEGLECTED TROPICAL DISEASES
(2023)
Article
Microbiology
Sarah Schurig, Rea Kobialka, Andy Wende, Md Anik Ashfaq Khan, Phillip Luebcke, Elias Eger, Katharina Schaufler, Arwid Daugschies, Uwe Truyen, Ahmed Abd El Wahed
Summary: In this study, different nucleic acid extraction methods were tested and it was found that alkali, proteinase K, and magnetic beads were the most effective for extracting DNA from bacteria and protozoa. These methods showed high sensitivity and were able to detect small amounts of bacterial and protozoan samples in each reaction.