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Entamoeba histolytica, the Causative Agent of Amoebiasis
Published in Hajiya Mairo Inuwa, Ifeoma Maureen Ezeonu, Charles Oluwaseun Adetunji, Emmanuel Olufemi Ekundayo, Abubakar Gidado, Abdulrazak B. Ibrahim, Benjamin Ewa Ubi, Medical Biotechnology, Biopharmaceutics, Forensic Science and Bioinformatics, 2022
Charles Oluwaseun Adetunji, Oyetunde T. Oyeyemi
Molecular based methods have helped to differentiate between these species thus solving some of the sensitivity and specificity deficiencies attributed to the classical microscopy utilized for recognition of protozoan pathogens (Pechangou et al., 2015). Multiplex polymerase chain reaction (mPCR), restriction fragment length polymorphism (RFLP), real-time PCR (RT-PCR), and gene sequencing are among the molecular tools developed for the identification of Entamoeba species (Samie et al., 2008; Bruijnesteijn van Coppenraet et al., 2009). Other possibility such as the use of Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to characterize Entamoeba species-specific amino acids and proteins has been explored. This proteomics approach has been acclaimed to offer a better method of differentiating between Entamoeba spp. compared to molecular methods because it is less protracted and less costly (Calderaro et al., 2015).
Infectious Bursal Disease
Published in Moayad N. Khalaf, Michael Olegovich Smirnov, Porteen Kannan, A. K. Haghi, Environmental Technology and Engineering Techniques, 2020
Reverse transcriptase–polymerase chain reaction (RT-PCR) was a molecular tool frequently applied for IBD diagnosis. Wu et al. (1997) developed the first PCR test for the detection of IBDV. A set of primers that specified a 150 bp fragment in segment A of IBDV genome was used to distinguish the IBDV from other infections in chicks. The PCR could detect 2 fg of IBDV RNA. A sensitive and specific multiplex polymerase chain reaction (mPCR) was developed and optimized for simultaneous detection and differentiation of avian reovirus (ARV), avian adenovirus group I (AAV-I), infectious bursal disease virus (IBDV) and chicken anemia virus (CAV) (Caterina et al., 2004). Zhang et al. (2002) detected IBDV during early stages of the disease by in situ RT-PCR using primers specific for VP4 region of segment A of IBDV. Kataria et al. (2004) performed RT-PCR on 17 bursal samples suspected for IBD collected from different parts of India. Multiple alignments with different strains suggested that simultaneous mutations in different regions were the most probable cause for strain variations. It was not possible to predict the most important amino acid residues without complete knowledge of the three-dimensional structure of the viral proteins.
Extended local similarity analysis (eLSA) reveals unique associations between bacterial community structure and odor emission during pig carcasses decomposition
Published in Journal of Environmental Science and Health, Part A, 2018
Bo-Min Ki, Hee Wook Ryu, Kyung-Suk Cho
DNA was extracted using the NucleoSpin Soil kit (Macherey-Nagel GmbH, Düren, Germany), and quantified using an ASP-2680 spectrophotometer (ACTGene, Piscataway, NJ, USA).[21] Ion Torrent sequencing was performed in duplicate on DNA to analyze bacterial communities. Thus, there were 56 sequencing libraries (samples on days 0, 31, 45, 76, 127, 161 and 225 in duplicate) for each manufacturing process. For multiplex polymerase chain reaction (PCR), different composite primer sets were used, based on the 340F and 805R primer sets for bacteria.[22] The detailed process was described in a previous study.[22] The purified DNA was analyzed on an Ion Torrent PGM system (Life Technologies, Grand Island, USA) by Macrogen Incorporation (Seoul, South Korea).