THE MODIFICATION OF DNA ISOLATION METHOD TO INCREASE PCR SENSITIVITY IN LABORATORY DIAGNOSTICS OF INFECTIOUS DISEASES

Author(s):  
K.A. Osyanin ◽  
◽  
K.V. Usoltsev ◽  
D.A. Mirgazov ◽  
L.I. Zaynullin ◽  
...  
2021 ◽  
Author(s):  
Bruno Marçal Repolês ◽  
Choco Michael Gorospe ◽  
Phong Tran ◽  
Anna Karin Nilsson ◽  
Paulina H. Wanrooij

AbstractThe integrity of mitochondrial DNA (mtDNA) isolated from solid tissues is critical for analyses such as long-range PCR. We show that a commonly-used DNA isolation procedure preferentially introduces strand breaks into the mtDNA extracted from the skeletal muscle of aged mice, while mtDNA from adult animals is less affected. We present a comparison of mtDNA isolation methods and identify one that avoids this biased loss of muscle mtDNA integrity. Our results highlight the importance of a careful choice of mtDNA isolation method and serve as a resource to researchers planning analysis of mtDNA isolated from solid tissues.


Author(s):  
D. V. Utkin ◽  
N. A. Ossina ◽  
V. E. Kouklev ◽  
P. S. Erokhin ◽  
S. A. Scherbakova ◽  
...  

The review presents the functional characteristics of the main types of biosensors: electrochemical, piezoelectric and optical. Shown are the examples of biosensors application for pathogenic biological agents detection. The prospects of biosensors development for laboratory diagnostics of particularly dangerous infectious diseases are discussed.


1999 ◽  
Vol 65 (6) ◽  
pp. 2307-2311 ◽  
Author(s):  
R. R. de Moraes ◽  
J. E. Maruniak ◽  
J. E. Funderburk

ABSTRACT Two methods, phenol-ether and magnetic capture-hybridization (MCH), were developed and compared with regard to their sensitivities and abilities to extract the DNA of the insect baculovirus Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) from soil and to produce DNA amplifiable by PCR. Laboratory experiments were performed with 0.25 g of autoclaved soil inoculated with different viral concentrations to optimize both methods of baculovirus DNA extraction and to determine their sensitivities. Both procedures produced amplifiable DNA; however, the MCH method was 100-fold more sensitive than the phenol-ether procedure. The removal of PCR inhibitors from the soil appeared to be complete when MCH was used as the viral DNA isolation method, because undiluted aliquots of the DNA preparations could be amplified by PCR. The phenol-ether procedure probably did not completely remove PCR inhibitors from the soil, since PCR products were observed only when the AgMNPV DNA preparations were diluted 10- or 100-fold. AgMNPV DNA was detected in field-collected soil samples from 15 to 180 days after virus application when the MCH procedure to isolate DNA was coupled with PCR amplification of the polyhedrin region.


1999 ◽  
Vol 9 (2) ◽  
pp. 111-114 ◽  
Author(s):  
Benjamin Kaufman ◽  
Stacy Richards ◽  
David A Dierig

2011 ◽  
Vol 86 (3) ◽  
pp. 310-312 ◽  
Author(s):  
Jeff A. Brady ◽  
Jennifer B. Faske ◽  
Jessica M. Castañeda-Gill ◽  
Jonathan L. King ◽  
Forrest L. Mitchell

2014 ◽  
Vol 84 (1) ◽  
pp. 267-272 ◽  
Author(s):  
J. R. Alvarado Bremer ◽  
B. L. Smith ◽  
D. L. Moulton ◽  
C.-P. Lu ◽  
M. Cornic

2021 ◽  
Vol 20 (4) ◽  
pp. 22-27
Author(s):  
O. V. Molochkova ◽  
O. B. Kovalev ◽  
O. V. Shamsheva ◽  
A. A. Sakharova ◽  
N. V. Sokolova ◽  
...  

The range of differential diagnostic search in the development of hemorrhagic colitis (hemocolitis) is wide enough and includes infectious and non-infectious factors. Purpose: clinical, laboratory and etiological analysis of bacterial diarrhea occurring with hemorrhagic colitis in the infectious diseases department.Materials and methods: a retrospective study of 141 case histories of those hospitalized in the infectious diseases department of the2 Children's City Clinical Hospital No. 9 named after G.N. Speransky of Moscow in 2019—2021 patients with clinical picture of hemocolitis. Hemocolitis was determined on the basis of macroscopic and microscopic signs (in scatology — mucus, leukocytes, erythrocytes) signs. All patients underwent routine laboratory examinations. The etiology was verified using modern methods of laboratory diagnostics (bacteriological analysis of feces, Latex test, ICA, ELISA, PCR, IHR).Results. There were 137 patients with infectious hemocolitis. Inflammatory bowel disease was diagnosed in 3 children, anus fissure — in 1 child. Young children under 3 years of age prevailed (77%). The etiology of infectious hemocolitis was deciphered in 47 patients (34%). Salmonellosis (36%) and shigellosis (36%) prevailed. Campylobacteriosis, clostridiosis-dificile and klebsiellosis accounted for 11%, 9% and 6% of cases, respectively. Yersiniosis was detected in 1 child at the age of 5 months. The severe form was found in 5.8% of cases, in most cases with shigellosis. Symptoms of intoxication and febrile fever were expressed in all patients, vomiting — in 28.5%, abdominal pain — in 94%, mesenteric adenitis on ultrasound — in 15%, diarrhea with a frequency of more than 5 times a day — in 84%, dehydration — in 64%, intercurrent diseases (ARVI, pneumonia) — in 41.3% of cases. Inflammatory changes in infectious hemocolitis were manifested by an increase in C-reactive protein in 71% (23.91 ± 24.17 mg/l), leukocytosis — in 69% (11.58 ± 3.52 х103 / μl), thrombocytosis — in 26%, an increase in the relative number of stab neutrophils in the general blood test in 78% of cases (10.95 ± 0.4%).Conclusions. Differential diagnostic search in the development of hemocolitis should include modern diagnostic methods, if necessary, additional instrumental studies and specialist consultations to exclude inflammatory bowel diseases.


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