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"loop-mediated isothermal amplification"

Brief Communications

Loop-Mediated Isothermal Amplification Targeting Actin DNA of Trichomonas vaginalis
Youn-Kyoung Goo, Won-Sik Shin, Hye-Won Yang, So-Young Joo, Su-Min Song, Jae-Sook Ryu, Hyun-Hee Kong, Won-Ki Lee, Dong-Il Chung, Yeonchul Hong
Korean J Parasitol 2016;54(3):329-334.
Published online June 30, 2016
DOI: https://doi.org/10.3347/kjp.2016.54.3.329
Trichomoniasis caused by Trichomonas vaginalis is a common sexually transmitted disease. Its association with several health problems, including preterm birth, pelvic inflammatory disease, cervical cancer, and transmission of human immunodeficiency virus, emphasizes the importance of improved access to early and accurate detection of T. vaginalis. In this study, a rapid and efficient loop-mediated isothermal amplification-based method for the detection of T. vaginalis was developed and validated, using vaginal swab specimens from subjects suspected to have trichomoniasis. The LAMP assay targeting the actin gene was highly sensitive with detection limits of 1 trichomonad and 1 pg of T. vaginalis DNA per reaction, and specifically amplified the target gene only from T. vaginalis. Validation of this assay showed that it had the highest sensitivity and better agreement with PCR (used as the gold standard) compared to microscopy and multiplex PCR. This study showed that the LAMP assay, targeting the actin gene, could be used to diagnose early infections of T. vaginalis. Thus, we have provided an alternative molecular diagnostic tool and a point-of-care test that may help to prevent trichomoniasis transmission and associated complications.

Citations

Citations to this article as recorded by  Crossref logo
  • Loop‐Mediated Isothermal Amplification (LAMP) for the Diagnosis of Sexually Transmitted Infections: A Review
    Yasaman Ahmadi, Yejiong Yu, Zhanfeng Cui, Wei E. Huang, Monique I. Andersson
    Microbial Biotechnology.2025;[Epub]     CrossRef
  • A novel detection method based on MIRA-CRISPR/Cas13a-LFD targeting the repeated DNA sequence of Trichomonas vaginalis
    Zhenke Yang, Jinghui Wang, Yiming Qi, Yiping Shi, Fakun Li, Weijuan Wang, Xiaowei Tian, Xuefang Mei, Zhenchao Zhang, Shuai Wang
    Parasites & Vectors.2024;[Epub]     CrossRef
  • A novel fluoro colorimetric Loop mediated isothermal amplification (LAMP) assay for detection of Trichomonas vaginalis
    Shoorashetty Manohar Rudresh, Pareyam Pooja, Pattacheravanda Nanaiah Shakuntala, Kanta Madhu
    Indian Journal of Medical Microbiology.2024; 49: 100610.     CrossRef
  • Establishment of a programmatic detection method for Trichomonas vaginalis based on double antibody sandwich ELISA targeting TvCP39 antigen
    Yuhua Li, Fakun Li, Wenjie Tian, Yani Zhang, Weijuan Wang, Zhenke Yang, Xiaowei Tian, Shuai Wang, Xuefang Mei, Zhenchao Zhang
    Acta Tropica.2024; 260: 107489.     CrossRef
  • Label-free electrochemical DNA biosensing of MR TV 29 18s ribosomal RNA gene of Trichomonas vaginalis by signalization of non-spherical gold nanoparticles
    R. Dehdari Vais, H. Heli, N. Sattarahmady
    Materials Today Communications.2023; 34: 105123.     CrossRef
  • Construction a novel detection method for Trichomonas vaginalis based on recombinant enzyme polymerase amplification targeting the Actin gene
    Fakun Li, Yangyang Deng, Wanxin Sheng, Xihui Gao, Weijuan Wang, Zhili Chu, Xuefang Mei, Zhenke Yang, Xiaowei Tian, Shuai Wang, Zhenchao Zhang
    Journal of Eukaryotic Microbiology.2023;[Epub]     CrossRef
  • A novel and ultrasensitive label-free electrochemical DNA biosensor for Trichomonas vaginalis detection based on a nanostructured film of poly(ortho-aminophenol)
    Rezvan Dehdari Vais, Hossein Heli, Naghmeh Sattarahmady, Afshin Barazesh
    Synthetic Metals.2022; 287: 117082.     CrossRef
  • Omics Analyses of Trichomonas vaginalis Actin and Tubulin and Their Participation in Intercellular Interactions and Cytokinesis
    Sebastián Lorenzo-Benito, Luis Alberto Rivera-Rivas, Lizbeth Sánchez-Ayala, Jaime Ortega-López, Octavio Montes-Flores, Daniel Talamás-Lara, Rossana Arroyo
    Genes.2022; 13(6): 1067.     CrossRef
  • Photo-genosensor for Trichomonas vaginalis based on gold nanoparticles-genomic DNA
    S. Ilbeigi, R. Dehdari Vais, N. Sattarahmady
    Photodiagnosis and Photodynamic Therapy.2021; 34: 102290.     CrossRef
  • Loop mediated isothermal amplification assay for detection of Trichomonas vaginalis in vaginal swabs among symptomatic women from North India
    S. Khurana, R. Dadwal, N. Sharma, A. Mewara, S. Singh, R. Bagga, R. Yadav, S. Sethi
    Letters in Applied Microbiology.2020; 70(3): 196.     CrossRef
  • Establishment and application of isothermal amplification techniques for the detection of heat-stable I enterotoxin of enterotoxigenic Escherichia coli
    Junjun Zhai, Zhang Yan, Feng Ping, Qu Lei, Xuelong Chen, Yanping Qi, Tianwen Wang
    PLOS ONE.2020; 15(4): e0230881.     CrossRef
  • Development of a convenient detection method for Trichomonas vaginalis based on loop-mediated isothermal amplification targeting adhesion protein 65
    Yuhua Li, Shuai Wang, Haoran Li, Xiaoxiao Song, Hao Zhang, Yujuan Duan, Chengyang Luo, Bingli Wang, Sifan Ji, Qing Xie, Zhenchao Zhang
    BMC Infectious Diseases.2020;[Epub]     CrossRef
  • Label-free ultrasensitive electrochemical genosensing of Trichomonas vaginalis using anisotropic-shaped gold nanoparticles as a platform, a repeated sequence of the parasite DNA as a probe, and toluidine blue as a redox marker
    N. Delshadi-Jahromi, R. Nazari-Vanani, H. Yadegari, N. Sattarahmady, G.R. Hatam, H. Heli
    Sensors and Actuators B: Chemical.2018; 273: 234.     CrossRef
  • Real-time loop-mediated isothermal amplification for rapid detection of Enterocytozoon hepatopenaei
    Shao-Xin Cai, Fan-De Kong, Shu-Fei Xu, Cui-Luan Yao
    PeerJ.2018; 6: e5993.     CrossRef
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Detection of Acute Toxoplasmosis in Pigs Using Loop-Mediated Isothermal Amplification and Quantitative PCR
Yanhua Wang, Guangxiang Wang, Delin Zhang, Hong Yin, Meng Wang
Korean J Parasitol 2013;51(5):573-577.
Published online October 31, 2013
DOI: https://doi.org/10.3347/kjp.2013.51.5.573

A loop-mediated isothermal amplification (LAMP) assay allows rapid diagnosis of Toxoplasma gondii infection. In the present study, the LAMP assay was evaluated using blood from both naturally and experimentally infected pigs. The sensitivity of the LAMP assay was compared with that of Q-PCR. Both assays detected T. gondii in the blood of experimentally infected pigs, with 100% agreement. In infected blood samples, the parasite was detected as early as 2 days post-infection and reached a peak in 3-5 days. In 216 field serum samples, the detection rates of LAMP and Q-PCR assays were 6.9% and 7.8%, respectively. This result indicates that the sensitivity of the LAMP assay was slightly lower than that of the Q-PCR assay. However, the LAMP may be an attractive diagnostic method in conditions where sophisticated and expensive equipment is unavailable. This assay could be a powerful supplement to current diagnostic methods.

Citations

Citations to this article as recorded by  Crossref logo
  • First report of molecular detection and phylogenetic analysis of Toxoplasma Gondii in soil, water and vegetables from Chandigarh city, India
    Divya Rattan, Priya Datta, Devyani Sharma, Chandra Kanta Bhusal, Rakesh Sehgal
    Scientific Reports.2025;[Epub]     CrossRef
  • Detection of chronic toxoplasmosis in the brain of mice using loop-mediated isothermal amplification (LAMP) and conventional PCR
    Mona K. Hegazy, Nora E. Saleh, Wafaa A. Aboukamar
    Experimental Parasitology.2023; 251: 108556.     CrossRef
  • Immune system roles in pathogenesis, prognosis, control, and treatment of Toxoplasma gondii infection
    Mohammad Mahdi Jafari, Zahra Azimzadeh Tabrizi, Mohammad Saaid Dayer, Nazanin Atieh Kazemi-Sefat, Mahshid Mohtashamifard, Rahimeh Mohseni, Atefeh Bagheri, Saeed Bahadory, Amir Karimipour-Saryazdi, Fatemeh Ghaffarifar
    International Immunopharmacology.2023; 124: 110872.     CrossRef
  • Clinical validation of visual LAMP and qLAMP assays for the rapid detection of Toxoplasma gondii
    Zhi Cao, Ke Zhang, Dehua Yin, Qiaoya Zhang, Ying Yu, Jianxin Wen, Hongbo Ni
    Frontiers in Cellular and Infection Microbiology.2022;[Epub]     CrossRef
  • RAA-Cas12a-Tg: A Nucleic Acid Detection System for Toxoplasma gondii Based on CRISPR-Cas12a Combined with Recombinase-Aided Amplification (RAA)
    Qiao-Ni Ma, Meng Wang, Lai-Bao Zheng, Zi-Qin Lin, Muhammad Ehsan, Xing-Xing Xiao, Xing-Quan Zhu
    Microorganisms.2021; 9(8): 1644.     CrossRef
  • Loop mediated isothermal amplification (LAMP) of Toxoplasma DNA from dried blood spots
    Mona K. Hegazy, Soha I. Awad, Nora E. Saleh, Mamdouh M. Hegazy
    Experimental Parasitology.2020; 211: 107869.     CrossRef
  • Advances in serological, imaging techniques and molecular diagnosis of Toxoplasma gondii infection
    Ali Rostami, Panagiotis Karanis, Shirzad Fallahi
    Infection.2018; 46(3): 303.     CrossRef
  • Molecular detection and phylogenetic analyses ofToxoplasma gondiifrom naturally infected sheep in Northern and Central Tunisia
    Mariem Rouatbi, Yosra Amdouni, Safa Amairia, Mohamed R. Rjeibi, Said Sammoudi, Mourad Rekik, Mohamed Gharbi
    Veterinary Medicine and Science.2017; 3(1): 22.     CrossRef
  • Comparison of PCR assays targeting the multi-copy targets B1 gene and 529 bp repetitive element for detection of Toxoplasma gondii in swine muscle
    Fabrizia Veronesi, Azzurra Santoro, Giovanni Luigi Milardi, Manuela Diaferia, Raffaella Branciari, Dino Miraglia, Attilia Cioffi, Simona Gabrielli, David Ranucci
    Food Microbiology.2017; 63: 213.     CrossRef
  • Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran
    Amir Abdoli, Abdolhossein Dalimi, Haleh Soltanghoraee, Fatemeh Ghaffarifar
    Journal of Parasitic Diseases.2016; 40(4): 1317.     CrossRef
  • Development and application of loop-mediated isothermal amplification assays based on ITS-1 for rapid detection of Toxoplasma gondii in pork
    Xunhui Zhuo, Bin Huang, Jiaqing Luo, Haijie Yu, Baolong Yan, Yi Yang, Aifang Du
    Veterinary Parasitology.2015; 208(3-4): 246.     CrossRef
  • Diagnosis of toxoplasmosis and typing of Toxoplasma gondii
    Quan Liu, Ze-Dong Wang, Si-Yang Huang, Xing-Quan Zhu
    Parasites & Vectors.2015;[Epub]     CrossRef
  • Loop-Mediated Isothermal Amplification for Rapid and Semiquantitative Detection of Loa loa Infection
    Papa M. Drame, Doran L. Fink, Joseph Kamgno, Jesica A. Herrick, Thomas B. Nutman, M. J. Loeffelholz
    Journal of Clinical Microbiology.2014; 52(6): 2071.     CrossRef
  • 7,576 View
  • 94 Download
  • Crossref

Original Article

Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Acanthamoeba
Hye-Won Yang, Yu-Ran Lee, Noboru Inoue, Bijay Kumar Jha, Dinzouna-Boutamba Sylvatrie Danne, Hong-Kyun Kim, Junhun Lee, Youn-Kyoung Goo, Hyun-Hee Kong, Dong-Il Chung, Yeonchul Hong
Korean J Parasitol 2013;51(3):269-277.
Published online June 30, 2013
DOI: https://doi.org/10.3347/kjp.2013.51.3.269

Amoebic keratitis (AK) caused by Acanthamoeba is one of the most serious corneal infections. AK is frequently misdiagnosed initially as viral, bacterial, or fungal keratitis, thus ensuring treatment delays. Accordingly, the early detection of Acanthamoeba would contribute significantly to disease management and selection of an appropriate anti-amoebic therapy. Recently, the loop-mediated isothermal amplification (LAMP) method has been applied to the clinical diagnosis of a range of infectious diseases. Here, we describe a rapid and efficient LAMP-based method targeting Acanthamoeba 18S rDNA gene for the detection of Acanthamoeba using clinical ocular specimens in the diagnosis of AK. Acanthamoeba LAMP assays detected 11 different strains including all AK-associated species. The copy number detection limit for a positive signal was 10 DNA copies of 18S rDNA per reaction. No cross-reactivity with the DNA of fungi or other protozoa was observed. The sensitivity of LAMP assay was higher than those of Nelson primer PCR and JDP primer PCR. In the present study, LAMP assay based on directly heat-treated samples was found to be as efficient at detecting Acanthamoeba as DNA extracted using a commercial kit, whereas PCR was only effective when commercial kit-extracted DNA was used. This study showed that the devised Acanthamoeba LAMP assay could be used to diagnose AK in a simple, sensitive, and specific manner.

Citations

Citations to this article as recorded by  Crossref logo
  • Ultrasensitive and rapid diagnostic tool for detection of Acanthamoeba castellanii
    Susanna Haapanen, Maarit S. Patrikainen, Seppo Parkkila
    Diagnostic Microbiology and Infectious Disease.2023; 107(2): 116014.     CrossRef
  • A simple and visible detection method for the rapid diagnosis of Ustilaginoidea virens in rice seeds by a loop‐mediated isothermal amplification assay
    Wei Wang, Hang Yin, Ning Huang, Cuijing Zhu, Yufei Wang, Xintong Qi, Lu Ma, Yunxin Fan, Yao Yu, Hongsheng Zhang, Yongmei Bao
    Journal of Phytopathology.2021; 169(6): 369.     CrossRef
  • Efficient nested-PCR-based method development for detection and genotype identification of Acanthamoeba from a small volume of aquatic environmental sample
    Tsui-Kang Hsu, Jung-Sheng Chen, Hsin-Chi Tsai, Chi-Wei Tao, Yu-Yin Yang, Ying-Chin Tseng, Yi-Jie Kuo, Dar-Der Ji, Jagat Rathod, Bing-Mu Hsu
    Scientific Reports.2021;[Epub]     CrossRef
  • Development of Visually Improved Loop Mediated Isothermal Amplification for the Diagnosis of Plasmodium vivax Malaria in a Tertiary Hospital in Chandigarh, North India
    Hargobinder Kaur, Rakesh Sehgal, Devendra Bansal, Ali A. Sultan, Ashish Bhalla, Sunit C. Singhi
    The American Journal of Tropical Medicine and Hygiene.2018; 98(5): 1374.     CrossRef
  • Detection of Acanthamoeba spp. in water samples collected from natural water reservoirs, sewages, and pharmaceutical factory drains using LAMP and PCR in China
    Anna Lass, Milena Guerrero, Xiuping Li, Gabriele Karanis, Liqing Ma, Panagiotis Karanis
    Science of The Total Environment.2017; 584-585: 489.     CrossRef
  • Water-borne protozoa parasites: The Latin American perspective
    Félix Manuel Rosado-García, Milena Guerrero-Flórez, Gabriele Karanis, María Del Carmen Hinojosa, Panagiotis Karanis
    International Journal of Hygiene and Environmental Health.2017; 220(5): 783.     CrossRef
  • Evaluation of Loop-mediated Isothermal Amplification Assay for Rapid Diagnosis of Acanthamoeba Keratitis
    Abhishek Mewara, Sumeeta Khurana, Shakila Yoonus, Kirti Megha, Parveen Tanwar, Amit Gupta, Rakesh Sehgal
    Indian Journal of Medical Microbiology.2017; 35(1): 90.     CrossRef
  • Acanthamoeba keratitis: improving the Scottish diagnostic service for the rapid molecular detection of Acanthamoeba species
    Claire Low Alexander, Michael Coyne, Brian Jones, Deepa Anijeet
    Journal of Medical Microbiology .2015; 64(7): 682.     CrossRef
  • Molecular diagnosis in clinical parasitology: When and why?
    Samson SY Wong, Kitty SC Fung, Sandy Chau, Rosana WS Poon, Sally CY Wong, Kwok-Yung Yuen
    Experimental Biology and Medicine.2014; 239(11): 1443.     CrossRef
  • 11,275 View
  • 96 Download
  • Crossref
Brief Communication
Sensitive and Rapid Detection of Giardia lamblia Infection in Pet Dogs using Loop-Mediated Isothermal Amplification
Jie Li, Peiyuan Wang, Aiguo Zhang, Ping Zhang, Muhamd Alsarakibi, Guoqing Li
Korean J Parasitol 2013;51(2):237-241.
Published online April 25, 2013
DOI: https://doi.org/10.3347/kjp.2013.51.2.237

Giardia lamblia is recognized as one of the most prevalent parasites in dogs. The present study aimed to establish a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of G. lamblia from dogs. The fecal samples were collected and prepared for microscopic analysis, and then the genomic DNA was extracted directly from purified cysts. The concentration of DNA samples of G. lamblia were diluted by 10-fold serially ranging from 10-1 to 10-5 ng/?l for LAMP and PCR assays. The LAMP assay allows the amplification to be finished within 60 min under isothermal conditions of 63℃ by employing 6 oligonucleotide primers designed based on G. lamblia elongation factor 1 alpha (EF1α) gene sequence. Our tests showed that the specific amplification products were obtained only with G. lamblia, while no amplification products were detected with DNA of other related protozoans. Sensitivity evaluation indicated that the LAMP assay was sensitive 10 times more than PCR. It is concluded that LAMP is a rapid, highly sensitive and specific DNA amplification technique for detection of G. lamblia, which has implications for effective control and prevention of giardiasis.

Citations

Citations to this article as recorded by  Crossref logo
  • Evaluation of loop mediated isothermal amplification, quantitative real-time PCR, conventional PCR methods for identifying Ascaris lumbricoides in human stool samples
    Mamta Thakur, Abhishek Mewara, PVM Lakshmi, Sucheta Guleria, Sumeeta Khurana
    Diagnostic Microbiology and Infectious Disease.2025; 112(3): 116808.     CrossRef
  • Evaluation of Touchdown Loop-Mediated Isothermal Amplification for the Detection of Giardia duodenalis
    Márton Hartdégen, András József Laki, Kolos Farkasvölgyi, Kristóf Iván, Judit Plutzer
    Parasitologia.2025; 5(2): 25.     CrossRef
  • Real‐time loop‐mediated isothermal amplification (real‐time LAMP) assay for rapid diagnosis of amoebic liver abscess
    Sandhya Khunger, Abhishek Mewara, Upninder Kaur, Ajay Duseja, Pallab Ray, Naveen Kalra, Navneet Sharma, Rakesh Sehgal
    Tropical Medicine & International Health.2024; 29(2): 104.     CrossRef
  • A colorimetric assay for Neospora caninum utilizing the loop-mediated isothermal amplification technique
    Tingting Liu, Kairao Hu, Meiyi Chen, Hongrong Hong, Xi Jiang, Rongsheng Huang, Yiwen Wang, Jing Huang, Xingang Yu, Quan Liu, Zhengkai Wei
    Research in Veterinary Science.2024; 179: 105395.     CrossRef
  • Advances in point-of-care and molecular techniques to detect waterborne pathogens
    Meysam Khodaparast, Dave Sharley, Stephen Marshall, Travis Beddoe
    npj Clean Water.2024;[Epub]     CrossRef
  • Detection of chronic toxoplasmosis in the brain of mice using loop-mediated isothermal amplification (LAMP) and conventional PCR
    Mona K. Hegazy, Nora E. Saleh, Wafaa A. Aboukamar
    Experimental Parasitology.2023; 251: 108556.     CrossRef
  • A rapid multiplex loop-mediated isothermal amplification (mLAMP) assay for detection of Entamoeba histolytica and Giardia duodenalis
    Abhishek Mewara, Sandhya Khunger, Chayan Sharma, Sivanantham Krishnamoorthi, Shreya Singh, Rakesh Yadav, Sumeeta Khurana, Rakesh Sehgal
    Letters in Applied Microbiology.2023;[Epub]     CrossRef
  • Disease Biomarkers of Giardiasis
    Norhamizah Roshidi, Norsyahida Arifin, Francisco Gonzalez Salazar
    Journal of Parasitology Research.2022; 2022: 1.     CrossRef
  • Optimization and validation of a loop-mediated isothermal amplification (LAMP) assay for detection of Giardia duodenalis in leafy greens
    Laura F. Lalonde, Vincent Xie, Jenna R. Oakley, Vladislav A. Lobanov
    Food and Waterborne Parasitology.2021; : e00123.     CrossRef
  • Loop mediated isothermal amplification (LAMP) of Toxoplasma DNA from dried blood spots
    Mona K. Hegazy, Soha I. Awad, Nora E. Saleh, Mamdouh M. Hegazy
    Experimental Parasitology.2020; 211: 107869.     CrossRef
  • Sensitive and rapid detection of Ortleppascaris sinensis (Nematoda: Ascaridoidea) by loop-mediated isothermal amplification
    Jinhong Zhao, Wei Xu, Genjun Tu, Yongkang Zhou, Xiaobing Wu
    PeerJ.2019; 7: e7607.     CrossRef
  • Development of a loop-mediated isothermal amplification technique and comparison with quantitative real-time PCR for the rapid visual detection of canine neosporosis
    Aongart Mahittikorn, Nipa Thammasonthijarern, Amonrattana Roobthaisong, Ruenruetai Udonsom, Supaluk Popruk, Sukhontha Siri, Hirotake Mori, Yaowalark Sukthana
    Parasites & Vectors.2017;[Epub]     CrossRef
  • Development of a Loop Mediated Isothermal Amplification for Diagnosis ofAscaris lumbricoidesin Fecal Samples
    Esther A. Shiraho, Agola L. Eric, Ibrahim N. Mwangi, Geoffrey M. Maina, Joseph M. Kinuthia, Martin W. Mutuku, Robert M. Mugambi, Jackson M. Mwandi, Gerald M. Mkoji
    Journal of Parasitology Research.2016; 2016: 1.     CrossRef
  • Molecular Detection of <i>Giardia intestinalis</i> from Stray Dogs in Animal Shelters of Gyeongsangbuk-do (Province) and Daejeon, Korea
    Jin-Cheol Shin, Alisha Wehdnesday Bernardo Reyes, Sang-Hun Kim, Suk Kim, Hyung-Jin Park, Kyoung-Won Seo, Kun-Ho Song
    The Korean Journal of Parasitology.2015; 53(4): 477.     CrossRef
  • Loop-mediated isothermal amplification assay for detection ofHistomonas meleagridisinfection in chickens targeting the 18S rRNA sequences
    Jinjun Xu, Chanbao Qu, Jianping Tao
    Avian Pathology.2014; 43(1): 62.     CrossRef
  • Diagnostic testing for Giardia infections
    M. F. Heyworth
    Transactions of the Royal Society of Tropical Medicine and Hygiene.2014; 108(3): 123.     CrossRef
  • 10,467 View
  • 100 Download
  • Crossref