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Arifa Akram, S M Rashedul Islam, Saif Ullah Munshi, Shahina Tabassum
Transmission of Hepatitis B Virus (HBV) usually occurs due to the transfusion of blood or blood products from chronic HBV (CHB) or occult HBV infected (OBI) patients. Besides serological tests e.g. HBsAg and anti-HBc (total), detection of HBV-DNA is necessary for the diagnosis of OBI patients. Different nucleic acid tests (NATs) including real-time-Polymerase Chain Reaction (qPCR) are used for the detect HBV-DNA. The NATs are expensive and require technical expertise which are barriers to introducing them in resource-limited settings...
May 16, 2018: Journal of Viral Hepatitis
Panan Kanchanaphum
This study explores determining the sex of humans from blood stains taken from different surfaces and compares the time course of detection with the conventional PCR, Conventional Loop Mediated Isothermal Amplification (LAMP), and LAMP-Lateral Flow Dipstick (LFD). For the DNA templates, 7 male and 7 female blood stained samples were extracted and added to LAMP and PCR reaction solution to amplify the SRY gene. The DNA samples were extracted from the following blood stained materials: cloth, wood, clay, and tile...
2018: BioMed Research International
S U Perera, V Navaratne, A Nagahawatte, J Perera, C D Wijayarathna, J Alvitigala, C L Goonasekara
No abstract text is available yet for this article.
March 31, 2018: Ceylon Medical Journal
Hagen Frickmann, Rebecca Hinz, Sandra Rojak, Insa Bonow, Stefanie Ruben, Christine Wegner, Iris Zielke, Ralf Matthias Hagen, Egbert Tannich
BACKGROUND: We assessed a commercial loop-mediated amplification (LAMP) platform for its reliability as a screening tool for malaria parasite detection. METHODS: A total of 1000 blood samples from patients with suspected or confirmed malaria submitted to the German National Reference Center for Tropical Pathogens were subjected to LAMP using the Meridian illumigene Malaria platform. Results were compared with microscopy from thick and thin blood films in all cases...
May 12, 2018: Travel Medicine and Infectious Disease
Kazuya Shirato, Shohei Semba, Sherif A El-Kafrawy, Ahmed M Hassan, Ahmed M Tolah, Ikuyo Takayama, Tsutomu Kageyama, Tsugunori Notomi, Wataru Kamitani, Shutoku Matsuyama, Esam Ibraheem Azhar
Clinical detection of Middle East respiratory syndrome (MERS) coronavirus (MERS-CoV) in patients is achieved using genetic diagnostic methods, such as real-time RT-PCR assay. Previously, we developed a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for the detection of MERS-CoV [Virol J. 2014. 11:139]. Generally, amplification of RT-LAMP is monitored by the turbidity induced by precipitation of magnesium pyrophosphate with newly synthesized DNA. However, this mechanism cannot completely exclude the possibility of unexpected reactions...
May 12, 2018: Journal of Virological Methods
Saleh Alamer, Shimaa Eissa, Raja Chinnappan, Paul Herron, Mohammed Zourob
Cotton swab is the conventional swabbing tool that is usually applied for collecting pathogens from contaminated surfaces, followed by cells lysis and DNA extraction before subjecting to genetic analysis. However, such an approach is time consuming as it involves several steps and requires highly trained personnel to perform the experiment. In this study, we developed a new cotton swab-based detection system that involved integrating bacterial collection, preconcentration and detection on Q-tips. The platform is based on a sandwich assay that can detect different pathogens visually by color changes...
August 1, 2018: Talanta
Gamal Wareth, Mohamed El-Diasty, Falk Melzer, Jayaseelan Murugaiyan, Amir Abdulmawjood, Lisa D Sprague, Heinrich Neubauer
Trueperella pyogenes was isolated from a dog and a cat with a mixed infection with Brucella abortus . Both lived on a dairy cattle farm with a history of regular cases of abortion and mastitis. Identification of the bacteria was done by means of MALDI-TOF MS, loop-mediated isothermal amplification (LAMP) based on cpn 60, partial 16S rRNA sequencing, and growth on Loeffler Serum Medium. Isolation of Trueperella pyogenes on the dairy farm highlights its neglected role in reproduction failure and draws attention to its effects in the dairy industry in Egypt...
2018: Veterinary Medicine International
Madhvi Chahar, Anup Anvikar, Rajnikant Dixit, Neena Valecha
Loop mediated isothermal amplification (LAMP) assay is sensitive, prompt, high throughput and field deployable technique for nucleic acid amplification under isothermal conditions. In this study, we have developed and optimized four different visualization methods of loop-mediated isothermal amplification (LAMP) assay to detect Pfcrt K76T mutants of P. falciparum and compared their important features for one-pot in-field applications. Even though all the four tested LAMP methods could successfully detect K76T mutants of P...
May 8, 2018: Experimental Parasitology
Amal Elamin, Raouf Wahab Ali, Vidar Bakken
The microbial profile of aggressive periodontitis patients is considered to be complex with variations among populations in different geographical areas. The aim of this study was to assess the presences of 4 putative periodontopathic bacteria (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola) and 2 periodontal herpes viruses (Epstein-Barr virus type 1 [EBV-1] and human cytomegalovirus [HMCV]) in subgingival plaque of Sudanese subjects with aggressive periodontitis and healthy controls...
October 2017: Clinical and Experimental Dental Research
Agnese Cristini, Matthias Groh, Maiken S Kristiansen, Natalia Gromak
R-loops comprise an RNA/DNA hybrid and displaced single-stranded DNA. They play important biological roles and are implicated in pathology. Even so, proteins recognizing these structures are largely undefined. Using affinity purification with the S9.6 antibody coupled to mass spectrometry, we defined the RNA/DNA hybrid interactome in HeLa cells. This consists of known R-loop-associated factors SRSF1, FACT, and Top1, and yet uncharacterized interactors, including helicases, RNA processing, DNA repair, and chromatin factors...
May 8, 2018: Cell Reports
Xiao Huang, Xingyu Lin, Katharina Urmann, Lijie Li, Xing Xie, Sunny Jiang, Michael R Hoffmann
Model coliphages (e.g., ΦX174, MS2, and PRD1) have been widely used in place of pathogenic viruses to study their fate and transport in the environment and during wastewater treatment. Two groups of coliphages (F-specific and somatic) are being explored as indicators of viral fecal pollution in ambient water. However, the detection and quantification of coliphages still largely relies on time-consuming culture-based plaque assays. In this study, we developed an in-gel loop mediated isothermal amplification (gLAMP) system enabling coliphage MS2 quantification within 30 minutes using standard laboratory devices...
May 8, 2018: Environmental Science & Technology
Dong Eun Lee, Hanna Lee, Seon Duk Lee, Hyung Soo Han, Jae Young Choe, Sora Yun, Hong-Jun Park, Jungbae Park, Jong Kun Kim
Background: Nucleic acid amplification assays (NAAs), such as polymerase chain reaction or loop-mediated isothermal amplification (LAMP), are used for disease diagnosis. Current nucleic acid isolation kits require several hours for completion of protocol including the complicated handling steps. Objective: In this study, a simple and cost-effective nucleic acid preparation method was developed, and its performance was compared with those of commercial kits. Materials and Methods: RNA was prepared using our method and three commercial RNA isolation kits...
January 2018: Indian Journal of Medical Microbiology
Jiansen Gong, Linlin Zhuang, Di Zhang, Ping Zhang, Xinhong Dou, Chengming Wang
Antimicrobial resistance genes play an important role in mediating resistance to sulfonamide in Gram-negative bacteria. While PCR is the current method to detect sulfonamide resistance genes (sul1, sul2, sul3), it is time-consuming and costly and there is an urgent need to develop a more convenient, simpler and rapid test for the sul. In this study, we describe a multiplex loop-mediated isothermal amplification (m-LAMP) assay we developed for the rapid and simultaneous detection of three sul. This m-LAMP assay successfully detected seven reference strains with different sul genotypes, but was negative for nine sul-negative reference strains...
April 30, 2018: Foodborne Pathogens and Disease
Jussi Hiltunen, Christina Liedert, Marianne Hiltunen, Olli-Heikki Huttunen, Johanna Hiitola-Keinänen, Sanna Aikio, Mikko Harjanne, Marika Kurkinen, Leena Hakalahti, Luke P Lee
Microfluidic-based integrated molecular diagnostic systems, which are automated, sensitive, specific, user-friendly, robust, rapid, easy-to-use, and portable, can revolutionize future medicine. Current research and development largely relies on polydimethylsiloxane (PDMS) to fabricate microfluidic devices. Since the transition from the proof-of-principle phase to clinical studies requires a vast number of integrated microfluidic devices, there is a need for a high-volume manufacturing method of silicone-based microfluidics...
April 30, 2018: Lab on a Chip
Abu Naser Mohon, Didier Menard, Mohammad Shafiul Alam, Kevin Perera, Dylan R Pillai
Background: Artemisinin-resistant malaria (ARM) remains a significant threat to malaria elimination. In the Greater Mekong subregion, the prevalence of ARM in certain regions has reached greater than 90%. Artemisinin-resistant malaria is clinically identified by delayed parasite clearance and has been associated with mutations in the propeller domain of the kelch 13 gene. C580Y is the most prevalent mutation. The detection of ARM currently relies on labor-intensive and time-consuming methods such as clinical phenotyping or in vitro susceptibility testing...
April 2018: Open Forum Infectious Diseases
Michael S Reid, X Chris Le, Hongquan Zhang
Isothermal exponential amplification techniques, such as strand-displacement amplification (SDA), rolling circle amplification (RCA), loop-mediated isothermal amplification (LAMP), nucleic acid sequence-based amplification (NASBA), helicase-dependent amplification (HDA), and recombinase polymerase amplification (RPA), have great potential for on-site, point-of-care, and in-situ assay applications. These amplification techniques eliminate the need for temperature cycling required for polymerase chain reaction (PCR) while achieving comparable amplification yield...
April 27, 2018: Angewandte Chemie
Rahul Parashar, L D Singla, Kanisht Batra, Rajendra Kumar, Neeraj Kashyap, Paramjit Kaur, M S Bal
To unravel equid trypanosomosis caused by Trypanosoma evansi in Punjab state of India, a cross sectional study was designed by utilizing parasitological and sero-molecular tools with objective to assess the prevalence of T. evansi in association with various risk factors in all agroclimatic zones of Punjab state of India. Parasitological Romanowksy stained thin blood smears (RSTBS) to detect patent infection, molecular techniques polymerase chain reaction I (PCR I; TBR 1/2 primers; targeting minichromosomal satellite DNA of T...
April 23, 2018: Acta Tropica
C Manjunatha, Sapna Sharma, Deepika Kulshreshtha, Sangeeta Gupta, Kartar Singh, Subhash C Bhardwaj, Rashmi Aggarwal
Leaf rust of wheat caused by Puccinia triticina has significant impact on wheat production worldwide. Effective and quick detection methodologies are required to mitigate yield loss and time constraints associated with monitoring and management of leaf rust of wheat. In the present study, detection of P. triticina has been simplified by developing a rapid, reliable, efficient and visual colorimetric method i.e., loop mediated isothermal amplification of DNA (LAMP). Based on in silico analysis of P. triticina genome, PTS68, a simple sequence repeat was found highly specific to leaf rust fungus...
2018: PloS One
Ya Zhou, Jingfan Xiao, Xin Ma, Qiyao Wang, Yuanxing Zhang
In purpose of valid Streptococcus iniae detection, we established a colorimetric biosensor using gold nanoparticles (AuNPs) labeled with dual functional probes and along with loop-mediated isothermal amplification (LAMP) assay (LAMP-AuNPs). Based on the characteristics of self-aggregation and bio-conjugation with ligands, AuNPs were chosen for observable color change in tandem with LAMP amplification method to reach high sensitivity and easy operation. Meanwhile, the improvement of dual probes that could fully utilize the LAMP product gave the biosensor a stable result exhibition...
April 25, 2018: Applied Microbiology and Biotechnology
Tetsuya Satoh, Seiya Kouroki, Keita Ogawa, Yorika Tanaka, Kazutoshi Matsumura, Susumu Iwase
Identifying body fluids from forensic samples can provide valuable evidence for criminal investigations. Messenger RNA (mRNA)-based body fluid identification was recently developed, and highly sensitive parallel identification using reverse transcription polymerase chain reaction (RT-PCR) has been described. In this study, we developed reverse transcription loop-mediated isothermal amplification (RT-LAMP) as a simple, rapid assay for identifying three common forensic body fluids, namely blood, semen, and saliva, and evaluated its specificity and sensitivity...
April 25, 2018: Analytical and Bioanalytical Chemistry
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