A PCR Referenced Comparative Study for Evaluation of Different Mycobacterium tuberculosis DNA Extraction Methods Directly from Sputum and from LJ Culture Isolates in Egypt

2021 ◽  
Vol 30 (2) ◽  
pp. 59-65
Author(s):  
Heba Elshahawy ◽  
Ehab B. Rakha ◽  
Mona B. Elhadidi ◽  
Sanaa S. Hamam
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Dna Extraction ◽  
Solid Phase ◽  
Laboratory Diagnosis ◽  
Extraction Methods ◽  
Phase Method ◽  
Conventional Pcr ◽  
Dna Yield ◽  
Microbiological Methods ◽  
Solid Phase Method

Background: Tuberculosis is a critical infectious disease primarily affecting the lungs and is more common in developing countries. In the 21st century, it forms a significant problem for world public health especially with the emergence and rising of drug resistant TB. Microbiological methods are the clue for the laboratory diagnosis. The ordinary methods for TB identification showed either weak sensitivity as in microscopy or lateness for many weeks as in culture. The evolution in molecular biology gives a chance for fast diagnosis of Mycobacterium tuberculosis helping start proper treatment early and holding its spread. The initial critical step in PCR is DNA extraction. Objective: The aim to evaluate different extraction methods of Mycobacterium tuberculosis retrieved directly from sputum samples and from LJ isolates from same patients and comparing DNA yield using conventional PCR. Methodology: DNA from 32 sputum samples from TB patients extracted by solid, digestion and phenol extraction methods, DNA from 40 LJ isolates extracted by solid, boiling and Cetyl trimethylammonium bromide methods. Extracted DNA was evaluated by conventional PCR. Results: Among 32 sputum samples, the extracted DNA by phenol method was 21/32 (65.62%) with highest DNA yield, digestion method 14/32 (43.75%) and solid phase method 1/32 (2.5%) with least DNA yield. From 40 MTB LJ culture isolates, the extracted DNA by boiling method was 28/40 (70%), CTAB method 18/40 (45%) and solid phase method 2/40 (5%). Conclusion: Phenol method was the best method (mean rank 2.34) for DNA extraction from sputum samples, while the easy and economic boiling method was the best method (mean rank 2.45) for DNA extraction from LJ culture isolates. The worst method of DNA extraction from both sputum and culture was phase solid method. A greater and easier yield of DNA was obtained from MTB LJ Culture than sputum.

Peculiarities of a solid-phase method for the Al-Fe/SiO2 and Al-Co/SiO2 powder catalysts production

2019 ◽  
pp. 63-68
Author(s):  
V.A. Artyukh ◽  
◽  
V.N. Borsch ◽  
V.S. Yusupov ◽  
S.Ya. Zhuk ◽  
...  
Keyword(s):  
Solid Phase ◽  
Phase Method ◽  
Solid Phase Method ◽  
Sio2 Powder

Synthesis and biological activity of new metallocenic angiotensin II analogs

1988 ◽  
Vol 53 (11) ◽  
pp. 2914-2919 ◽  
Author(s):  
Pierrette Maes ◽  
Annie Ricouart ◽  
Emmanuel Escher ◽  
André Tartar ◽  
Christian Sergheraert
Keyword(s):  
Amino Acids ◽  
Biological Activity ◽  
Angiotensin Ii ◽  
Solid Phase ◽  
Rabbit Aorta ◽  
Phase Method ◽  
Solid Phase Method

Analogs of angiotensin II in which phenylalanine in position 8 was replaced with cymantrenylalanine or with its triphenylphosphine photosubstitution product were synthesized by the solid-phase method. On rabbit aorta strips, these peptides were found to be pure antagonists of angiotensin II. Their relative affinities are higher than most other analogs substituted in position 8 with bulky amino-acids.

Synthesis of four new V1 antagonists of arginine-vasopressin (AVP) containing new thioacids at position 1 and their vasoconstrictor activity towards isolated mesenteric arterial vessels of rats

1991 ◽  
Vol 56 (2) ◽  
pp. 491-498 ◽  
Author(s):  
Bernard Lammek ◽  
Izabela Derdowska ◽  
Tomasz M. Wierzba ◽  
Witold Juzwa
Keyword(s):  
Peptide Synthesis ◽  
Arginine Vasopressin ◽  
Solid Phase ◽  
Structural Features ◽  
Phase Method ◽  
Competitive Antagonist ◽  
Vasoconstrictor Effect ◽  
Solid Phase Method

In an attempt to determine some of the structural features in position 1 that account for V1 antagonism, four new analogues of arginine-vasopressin were synthesized and the effect of the modifications on the vasoconstrictor activity was checked using isolated mesenteric arterial vessels of rats. The protected precursors required for these analogues were synthesized by a solid phase method of peptide synthesis. One of the reported analogues, namely [1-(4-mercapto-4-tetrahydrothiopyraneacetic acid)., 2-O-methyltyrosine, 8-arginine]vasopressin appears to be a potent competitive antagonist of the vasoconstrictor effect by AVP.

Synthesis of a 21-Residue Fragment of Human Proinsulin by the Polyamide Solid Phase Method

1981 ◽  
Vol 362 (2) ◽  
pp. 833-840 ◽  
Author(s):  
Eric ATHERTON ◽  
Willy HÜBSCHER ◽  
Robert C. SHEPPARD ◽  
Vivienne WOOLLEY
Keyword(s):  
Solid Phase ◽  
Phase Method ◽  
Human Proinsulin ◽  
Solid Phase Method

scholarly journals Syntheses of two deacetyl-thymosin β4analogs and their anti-inflammatory effect on carrageenin-induced edema in the mouse paw

2001 ◽  
Vol 10 (2) ◽  
pp. 89-92 ◽  
Author(s):  
T. Abiko ◽  
R. Ogawa
Keyword(s):  
Marked Reduction ◽  
Solid Phase ◽  
The Other ◽  
Para Position ◽  
Phase Method ◽  
Antiinflammatory Effect ◽  
Solid Phase Method ◽  
Anti Inflammatory ◽  
Inflammatory Effect

Two {Met(0)6}deacetyl-thymosin β4analogs containing Phe(4F) or Tyr(Me) at position 12 were synthesized by the manual solid-phase method, and their anti-inflammatory effect on carrageenin-induced edema in the mouse paw was studied. Fluorination of the para-position of Phe12resulted in a marked antiinflammatory effect on carrageenin-induced edema in the mouse paw compared with that of our synthetic {Met(0)6}deacetyl-thymosin β4, but the other analog, {Met(0)6, Tyr(Me)12}deacetyl-thymosin β4, showed a marked reduction of the anti-inflammatory effect.

The Gas-Solid-Phase 2,5-Dioxopiperazine Synthesis. Cyclization of Vaporous Dipeptides on Silica Surface

1994 ◽  
Vol 59 (2) ◽  
pp. 461-466 ◽  
Author(s):  
Vladimir A. Basiuk ◽  
Taras Yu. Gromovoy
Keyword(s):  
Silica Surface ◽  
Solid Phase ◽  
Phase Method ◽  
Solid Phase Method

The "gas solid-phase" method is used for the preparation of both symmetric and asymmetric 2,5-dioxopiperazines via cyclization of vaporous linear dipeptides in the presence of silica.

Combustion Synthesis of La0.65Sr0.35MnO3 and its Sensing Properties for NO2

2016 ◽  
Vol 680 ◽  
pp. 208-211
Author(s):  
Lian Lian Wu ◽  
Qiang Li ◽  
Dan Yu Jiang ◽  
Jin Feng Xia
Keyword(s):  
Solid Phase ◽  
High Sensitivity ◽  
Combustion Method ◽  
Fast Response ◽  
Size Analysis ◽  
Phase Method ◽  
X Ray Diffraction ◽  
Ultrafine Structure ◽  
Nox Sensor ◽  
Solid Phase Method

In this paper, La0.65Sr0.35MnO3 (LSM) oxide powder with ultrafine structure has been synthesized by self-propagating combustion method. The powders were characterized by X-ray diffraction, scanning electron microscopy and laser size analysis. Compared to the powders prepared by traditional solid-phase method, the grain size of powders prepared by self-propagating combustion method is relatively small and uniform. Starting from ultrafine LSM powders, sensing electrode (SE) for NO2 mixed-potential sensors based on yttria-stablized zirconia (YSZ) was fabricated. As-obtained NO2 sensor displays fast response and high sensitivity (25.4mV/decade). The response values of the sensor have good linear relationship with the logarithm of NO2 concentration varying from 30ppm to 500ppm.Keywords:Self-propagating combustion method; La0.65Sr0.35MnO3; NOx sensor; YSZ

scholarly journals Assessing Quality and Functionality of DNA from Fresh and Archival Dried Blood Spots and Recommendations for Quality Control Guidelines

2007 ◽  
Vol 53 (8) ◽  
pp. 1401-1407 ◽  
Author(s):  
Malin Ida Linnea Sjöholm ◽  
Joakim Dillner ◽  
Joyce Carlson
Keyword(s):  
Dna Extraction ◽  
Multiple Displacement Amplification ◽  
Dried Blood Spots ◽  
Extraction Methods ◽  
Population Based ◽  
Alkaline Lysis ◽  
Screening Programs ◽  
Blood Spots ◽  
Dna Yield ◽  
Chelex 100

Abstract Background: Dried blood spots (DBS) are a convenient and inexpensive method for biobanking. Although many countries have established population-based DBS biobanks from neonatal screening programs, the quality and usefulness of DNA from DBS have not been extensively assessed. Methods: We compared 4 common DNA extraction methods (Qiagen, EZNA, Chelex 100, and alkaline lysis) in a pilot study using fresh DBS with known lymphocyte count. We assessed suitability for multiple displacement amplification (MDA) and subsequent single-nucleotide polymorphism (SNP) analyses. We selected the EZNA method for DNA extraction from archival samples up to 27 years old, stored at room temperature or −20 °C, and SNP analyses were performed after MDA. Results: Extraction using alkaline lysis failed in most tests, and Chelex 100 was unsuccessful in real-time PCR, whereas the EZNA and Qiagen methods were successful by all evaluated quality indices. DNA extraction by EZNA, MDA, and SNP analyses were successful for the archival samples stored at −20 °C. Conclusion: Routine protocols for evaluation of the quality and functional integrity of DNA based on DNA yield, DNA size, and quantification of amplifiable DNA allow use of sufficient template for MDA and successful SNP analyses from both primary DBS extract and MDA product. A single 3-mm disc can yield sufficient DNA for several thousand SNP analyses. DNA from DBS is thus suitable for genetic epidemiology studies.

Sign in / Sign up

Export Citation Format

Share Document