OPTIMASI DETEKSI GEN PADA Stelechocarpus burahol (Bl.) Hook.f. & Th. MENGGUNAKAN DIRECT KIR PCR

DNA isolation and purification in the conventional Polymerase Chain Reaction (PCR) process require reagents that are toxic, more costly and time consuming, and contamination. S. burahol leaves contain phenolics, flavonoids, and terpenoids which can interfere with DNA isolation. The use of direct PCR kits can detect genes without DNA extraction.The objective of study was to determinethe method of gene detection ofStelechocarpus buraholusing directPCRkit.In each location, one tree was taken as a source of leaf samples from Garut, Purwodadi Botanical Gardens, Kyai Langgeng Gardens, Yogyakarta Palace, Turi Sleman, Wanagama, Karanganyar, and South Kalimantan, except Bogor Botanical Gardens, two trees were taken. The primers used for the trials were ITS 1F primers and 4R primers. In the sequencing stage, PCR product samples of 40 -50 μl that showed positive results were detected by electrophoresis. The PCR product was measuredat ± 750 bp from ten samples. Direct PCR kits can be used for S. buraholgene detection, time and energy efficient, only requires a small amount of tissue, and reduces contamination due to DNA extraction. Direct PCR kits can be an effective method that can be utilized to detect target genesfor large populations.

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DNA Isolation of Microbial Contaminants in Aviation Turbine Fuel via Traditional Polymerase Chain Reaction (PCR) and Direct PCR. Preliminary Results

10.21236/ada446701 ◽

2005 ◽

Cited By ~ 4

Author(s):

Tracy R. Denaro ◽

Sarah K. Chelgren ◽

Jara N. Lang ◽

Ellen M. Strobel ◽

Lori M. T. Balster ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Dna Isolation ◽

Chain Reaction ◽

Direct Pcr ◽

Preliminary Results ◽

Polymerase Chain ◽

Microbial Contaminants ◽

Aviation Turbine Fuel

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INVESTIGATION OF ACCD3 GENE OF MYCOBACTERIUM TUBERCULOSIS IRAQI ISOLATES

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2018.v11i8.25269 ◽

2018 ◽

Vol 11 (8) ◽

pp. 208

Author(s):

Asra'a A Abdul Jalil ◽

Zahra M Al-khafaji ◽

Mushtak T Al-ouqaili

Keyword(s):

Mycobacterium Tuberculosis ◽

Drug Target ◽

Conventional Polymerase Chain Reaction ◽

Mycolic Acid ◽

Human Pathogens ◽

Mycolic Acids ◽

Pcr Product ◽

Malonyl Coa ◽

Polymerase Chain ◽

Acetyl Coenzyme A Carboxylase

Objective: Mycobacterium tuberculosis, one of the deadliest human pathogens, causes several million new infections and about 2 million fatalities annually. The cell wall of M. tuberculosis is endowed with a highly impermeable, complex array of diverse lipids such as mycolic acids, which bestow the bacterium with not only virulence but also resistance to host immunity and antibiotics.Methods: Mycobacterial lipid metabolism has thus emerged as an attractive target for the design and development of novel antimycobacterial therapeutics. The first committed step in the biosynthesis of mycolic acid is the carboxylation of acetyl-CoA to malonyl-CoA which is catalyzed by acetyl-coenzyme A carboxylase carboxyl transferase beta subunit (accD3), a primer pairs were designed computationally and used for the amplification of accD3 gene using conventional polymerase chain reaction (PCR) and sequencing the PCR product and analyze the results.Results: Two sequences of the detection gene (LprM gene) and eight sequences of accD3 gene under study were deposited at NCBI – GenBank database with accession numbers (LC009881, LC009880.1, LC006979, LC008196, LC009412, LC009414, LC034168, LC038020, LC041163, and LC041368) and primer pairs deposited at Probe database/NCBI with accession number Pr032816836.Conclusion: AccD3 gene is a good drug target in MDR M. tuberculosis strains.

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An unusual case of spinal cord restricted mycobacteriosis in a European mink

Tierärztliche Praxis Ausgabe K Kleintiere / Heimtiere ◽

10.1055/s-0038-1623679 ◽

2013 ◽

Vol 41 (01) ◽

pp. 63-66

Author(s):

D. Schaudien ◽

C. Flieshardt ◽

I. Moser ◽

H. Hotzel ◽

A. Tipold ◽

...

Keyword(s):

Spinal Cord ◽

Unusual Case ◽

Histological Evaluation ◽

Mustela Lutreola ◽

Chain Reaction ◽

European Mink ◽

Pcr Product ◽

Granulomatous Lesions ◽

The Central Nervous System ◽

Polymerase Chain

SummaryGranulomatous myelitis due to infection with Mycobacterium avium was diagnosed in a 4-year-old male neutered European mink (Mustela lutreola). The causative agent was detected by an acid-fast stain and further characterized by polymerase chain reaction and DNA sequencing of the PCR product. A thorough histological evaluation of the remaining organs revealed no granulomatous lesions or detectable acid-fast organisms. Although minks are generally highly susceptible for mycobacteria, localised infections, especially of the central nervous system, are unusual and may represent an atypical chronic form of the disease.

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Transmission of SARS-CoV-2 in Different Districts of a County in Istanbul, March to September 2020

Asia Pacific Journal of Public Health ◽

10.1177/10105395211009008 ◽

2021 ◽

pp. 101053952110090

Author(s):

Hüseyin Bilgin ◽

Ahmet Topuzoğlu ◽

Volkan Korten

Keyword(s):

Public Transportation ◽

Disease Transmission ◽

Geographical Information Systems ◽

Geographical Information ◽

Chain Reaction ◽

Heat Map ◽

Large Populations ◽

Polymerase Chain ◽

Education Tool ◽

Airborne Viruses

Epidemics caused by airborne viruses in cities with large populations create a big problem as in the current COVID-19 pandemic. Cramped lifestyle, busy workplaces, crowded public transportation, and higher household member counts are responsible for the transmission of the disease. In Turkey, Istanbul has taken the lead in the number of cases since the beginning of the epidemic. The excess population density is the major cause for disease transmission. It is essential to monitor the contaminated regions with geographical information systems on city maps. Outbreak maps visualize and help analyze the patterns of transmission and serve as a communication and education tool. A dynamic heat map video of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) polymerase chain reaction positive cases in a county of Istanbul was generated. The heat map visualizes how the epidemic spread to all the districts and the cumulative cases increased in one county of Istanbul with real attack rates.

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DMSO Improves the Ski-Slope Effect in Direct PCR

Applied Sciences ◽

10.3390/app11041943 ◽

2021 ◽

Vol 11 (4) ◽

pp. 1943

Author(s):

Joo-Young Kim ◽

Ju Yeon Jung ◽

Da-Hye Kim ◽

Seohyun Moon ◽

Won-Hae Lee ◽

...

Keyword(s):

Dna Sequences ◽

Pcr Amplification ◽

Analytical Techniques ◽

Direct Pcr ◽

Dna Profile ◽

Novel Technologies ◽

Specific Amplification ◽

Polymerase Chain ◽

Dna Profiles

Analytical techniques such as DNA profiling are widely used in various fields, including forensic science, and novel technologies such as direct polymerase chain reaction (PCR) amplification are continuously being developed in order to acquire DNA profiles efficiently. However, non-specific amplification may occur depending on the quality of the crime scene evidence and amplification methods employed. In particular, the ski-slope effect observed in direct PCR amplification has led to inaccurate interpretations of the DNA profile results. In this study, we aimed to reduce the ski-slope effect by using dimethyl sulfoxide (DMSO) in direct PCR. We confirmed that DMSO (3.75%, v/v) increased the amplification yield of large-sized DNA sequences more than that of small-sized ones. Using 50 Korean buccal samples, we further demonstrated that DMSO reduced the ski-slope effect in direct PCR. These results suggest that the experimental method developed in this study is suitable for direct PCR and may help to successfully obtain DNA profiles from various types of evidence at crime scenes.

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Granulocytic anaplasmosis in captive ring-tailed lemur (Lemur catta) in Poland

BMC Veterinary Research ◽

10.1186/s12917-021-02827-8 ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Łukasz Adaszek ◽

Anna Wilczyńska ◽

Jerzy Ziętek ◽

Marcin Kalinowski ◽

Oliwier Teodorowski ◽

...

Keyword(s):

Anaplasma Phagocytophilum ◽

Polymerase Chain Reaction Test ◽

Anaplasma Ovis ◽

Case Presentation ◽

Granulocytic Anaplasmosis ◽

Pcr Product ◽

Polymerase Chain ◽

Obligate Intracellular Bacteria ◽

Chain Reaction Test ◽

Anaplasma Bovis

Abstract Background Anaplasma are obligate intracellular bacteria and aetiological agents of tick-borne diseases of both veterinary and medical interest. The genus Anaplasma comprises six species: Anaplasma marginale, Anaplasma centrale, Anaplasma ovis, Anaplasma phagocytophilum, Anaplasma bovis and Anaplasma platys. They can infect humans, carnivores, ruminants, rodents, insectivores, birds and reptiles. The aim of this study was to present the first clinical case of granulocytic anaplasmosis in a captive ring-tailed lemur in Poland. Case presentation A 4-year-old female lemur presented anorexia, epistaxis and tick infestation. The microscopic examination of a blood smear revealed morulae in neutrophils. Polymerase chain reaction test and sequencing of obtained PCR product confirmed infection by the GU183908 Anaplasma phagocytophilum strain. Therapeutic protocol included doxycycline (2.5 mg/kg p.o., b.i.d.) for 3 weeks and the lemur recovered within 24 h. Conclusions This is the first report on granulocytic anaplasmosis in a ring-tailed lemur in Europe, indicating that A. phagocytophilum infection must also be considered in differential diagnosis in this animal species, especially in individuals with thrombocytopenia associated with Ixodes ricinus parasitism.

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