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[Application of multiplex-PCR for bifidobacteria and propionibacteria genus identification].

https://arctichealth.org/en/permalink/ahliterature262109
Source
Zh Mikrobiol Epidemiol Immunobiol. 2014 Sep-Oct;(5):78-82
Publication Type
Article
Author
S V Andriushchenko
N B Perunova
E V Ivanova
O V Bukharin
Source
Zh Mikrobiol Epidemiol Immunobiol. 2014 Sep-Oct;(5):78-82
Language
Russian
Publication Type
Article
Keywords
Bifidobacterium - classification - genetics - isolation & purification
DNA, Bacterial - genetics
Humans
Multiplex Polymerase Chain Reaction - methods
Propionibacterium - classification - genetics - isolation & purification
RNA, Ribosomal, 16S - genetics
Russia
Abstract
Creation of a PCR test-system for determination of Actinobacteria class bacteria belonging to 2 genera that are the most widely represented among obligate anaerobic microbiota of human intestine: Bifidobacterium and Propionibacterium.
8 strains of Bifidobacterium spp. and 6 strains of Propionibacterium genus were identified by morphologic, cultural and biochemical properties. Isolation of matrix DNA of the strains for PCR was carried out by "DNA-Express" kit (SPF "Lytech", Russia). Primers for determination of genus membership for obligate anaerobes were developed based on variability of 16S RNA gene by using "Lasergene 7.1" ("DNASTAR, Inc.", USA) program. PCR screening of the isolated DNA was carried out based on "Syntol" LLC primers and reagents. Amplicon detection was carried out by agarose gel electrophoresis.
Multiplexing of 2 different primer pairs in a single probe at.68 degrees C annealing temperature for 30 cycles showed the presence of non-specific amplicons that form in samples with bifidobacteria DNA-matrix. The increase of annealing temperature to 70 degrees C and reduction of the number of PCR cycles to 25 resulted in the exclusion of formation of non-specific amplicons. Because the annealing temperature reached the level of values optimal for Taq-polymerase, a 2-phase PCR algorithm could be implemented. This solution allowed reducing the overall time of reaction to 45 minutes. Further increase of annealing temperature to 72 degrees C and reduction of elongation phase up to 15 seconds at 30 PCR cycles did not result in a visible reduction of reaction effectiveness:
A rapid system for identification of Bifidobacterium and Pronionibacterium genera using a system of 2-phase multiple PCR was developed. The system is part of a screening system for identification of major genera and species of cultured obligate anaerobic bacteria isolated from human intestine biotopes.
PubMed ID
25536776 View in PubMed
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Asymptomatic carriers contribute to nosocomial Clostridium difficile infection: A cohort study of 4508 patients.

https://arctichealth.org/en/permalink/ahliterature283550
Source
Gastroenterology. 2017 Apr;152(5):1031-1041.e2
Publication Type
Article
Date
Apr-2017
Author
Thomas Blixt
Kim Oren Gradel
Christian Homann
Jakob Benedict Seidelin
Kristian Schønning
Anne Lester
Jette Houlind
Marie Stangerup
Magnus Gottlieb
Jenny Dahl Knudsen
Source
Gastroenterology. 2017 Apr;152(5):1031-1041.e2
Date
Apr-2017
Language
English
Publication Type
Article
Keywords
Adolescent
Adult
Aged
Aged, 80 and over
Asymptomatic Infections - epidemiology
Carrier State - diagnosis - epidemiology
Clostridium Infections - diagnosis - epidemiology
Clostridium difficile - genetics
Cohort Studies
Cross Infection - epidemiology
Denmark - epidemiology
Enterocolitis, Pseudomembranous - epidemiology
Female
Hospitalization
Hospitals, University
Humans
Male
Mass Screening
Middle Aged
Multiplex Polymerase Chain Reaction
Odds Ratio
Prospective Studies
Real-Time Polymerase Chain Reaction
Risk factors
Young Adult
Abstract
Nosocomial infections with Clostridium difficile present a considerable problem despite numerous attempts by health care workers to reduce risk of transmission. Asymptomatic carriers of C difficile can spread their infection to other patients. We investigated the effects of asymptomatic carriers on nosocomial C difficile infections.
We performed a population-based prospective cohort study at 2 university hospitals in Denmark, screening all patients for toxigenic C difficile in the intestine upon admittance, from October 1, 2012, to January 31, 2013. Screening results were blinded to patients, staff, and researchers. Patients were followed during their hospital stay by daily registration of wards and patient rooms. The primary outcomes were rate of C difficile infection in exposed and unexposed patients and factors associated with transmission.
C difficile infection was detected in 2.6% of patients not exposed to carriers and in 4.6% of patients exposed to asymptomatic carriers at the ward level (odds ratio for infection if exposed to carrier, 1.79; 95% confidence interval, 1.16-2.76). Amount of exposure correlated with risk of C difficile infection, from 2.2% in the lowest quartile to 4.2% in the highest quartile of exposed patients (P = .026). Combining the load of exposure to carriers and length of stay seemed to have an additive effect on the risk of contracting C difficile.
In a population-based prospective cohort study in Denmark, we found that asymptomatic carriers of toxigenic C difficile in hospitals increase risk of infection in other patients.
PubMed ID
28063955 View in PubMed
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Characterization of trh2 harbouring Vibrio parahaemolyticus strains isolated in Germany.

https://arctichealth.org/en/permalink/ahliterature270672
Source
PLoS One. 2015;10(3):e0118559
Publication Type
Article
Date
2015
Author
Silke Bechlars
Claudia Jäckel
Susanne Diescher
Doreen A Wüstenhagen
Stefan Kubick
Ralf Dieckmann
Eckhard Strauch
Source
PLoS One. 2015;10(3):e0118559
Date
2015
Language
English
Publication Type
Article
Keywords
Animals
Bacterial Proteins - chemical synthesis - genetics
Cell-Free System - chemistry
Erythrocytes - pathology
Fishes - microbiology
Genotype
Germany
Hemolysin Proteins - chemical synthesis - genetics
Hemolysis
Humans
Multiplex Polymerase Chain Reaction
Norway
Sheep - blood - microbiology
Vibrio parahaemolyticus - genetics - isolation & purification - pathogenicity
Virulence Factors - genetics
Abstract
Vibrio parahaemolyticus is a recognized human enteropathogen. Thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) as well as the type III secretion system 2 (T3SS2) are considered as major virulence factors. As tdh positive strains are not detected in coastal waters of Germany, we focused on the characterization of trh positive strains, which were isolated from mussels, seawater and patients in Germany.
Ten trh harbouring V. parahaemolyticus strains from Germany were compared to twenty-one trh positive strains from other countries. The complete trh sequences revealed clustering into three different types: trh1 and trh2 genes and a pseudogene ?trh. All German isolates possessed alleles of the trh2 gene. MLST analysis indicated a close relationship to Norwegian isolates suggesting that these strains belong to the autochthonous microflora of Northern Europe seawaters. Strains carrying the pseudogene ?trh were negative for T3SS2ß effector vopC. Transcription of trh and vopC genes was analyzed under different growth conditions. Trh2 gene expression was not altered by bile while trh1 genes were inducible. VopC could be induced by urea in trh2 bearing strains. Most trh1 carrying strains were hemolytic against sheep erythrocytes while all trh2 positive strains did not show any hemolytic activity. TRH variants were synthesized in a prokaryotic cell-free system and their hemolytic activity was analyzed. TRH1 was active against sheep erythrocytes while TRH2 variants were not active at all.
Our study reveals a high diversity among trh positive V. parahaemolyticus strains. The function of TRH2 hemolysins and the role of the pseudogene ?trh as pathogenicity factors are questionable. To assess the pathogenic potential of V. parahaemolyticus strains a differentiation of trh variants and the detection of T3SS2ß components like vopC would improve the V. parahaemolyticus diagnostics and could lead to a refinement of the risk assessment in food analyses and clinical diagnostics.
Notes
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Erratum In: PLoS One. 2015;10(4):e012656925901603
PubMed ID
25799574 View in PubMed
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Charcot-Marie-Tooth caused by a copy number variation in myelin protein zero.

https://arctichealth.org/en/permalink/ahliterature132705
Source
Eur J Med Genet. 2011 Nov-Dec;54(6):e580-3
Publication Type
Article
Author
Helle Høyer
Geir J Braathen
Anette K Eek
Camilla F Skjelbred
Michael B Russell
Author Affiliation
Section of Medical Genetics, Department of Laboratory Medicine, Telemark Hospital, Skien, Norway. helle.hoyer@sthf.no
Source
Eur J Med Genet. 2011 Nov-Dec;54(6):e580-3
Language
English
Publication Type
Article
Keywords
Adolescent
Adult
Age of Onset
Aged
Charcot-Marie-Tooth Disease - genetics
Child
Chromosome Breakpoints
Comparative Genomic Hybridization
DNA Copy Number Variations
Female
Genes, Dominant
Humans
Male
Middle Aged
Multiplex Polymerase Chain Reaction
Myelin P0 Protein - genetics
Norway
Pedigree
Peripheral Nervous System - pathology - physiopathology
Phenotype
Sequence Analysis, DNA
Abstract
Charcot-Marie-Tooth disease (CMT) is the most common inherited disorder of the peripheral nervous system. The majority has a duplication of the peripheral myelin protein 22. CMT is otherwise caused by point mutations or small insertions/deletions in one of the 44 known CMT genes.
Conventional sequencing of six CMT genes were followed by Multiplex Ligation-dependent Probe Amplification (MLPA), array Comparative Genomic Hybridization (aCGH) and breakpoint analysis in a large Norwegian CMT pedigree. Affected had an extra copy of the myelin protein zero (MPZ) gene.
To our knowledge this is the first non-peripheral myelin protein 22 copy number variation to cause Charcot-Marie-Tooth disease.
PubMed ID
21787890 View in PubMed
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Clinical evaluation of commercial nucleic acid amplification tests in patients with suspected sepsis.

https://arctichealth.org/en/permalink/ahliterature268237
Source
BMC Infect Dis. 2015;15:199
Publication Type
Article
Date
2015
Author
Lars Ljungström
Helena Enroth
Berndt E B Claesson
Ida Ovemyr
Jesper Karlsson
Berit Fröberg
Anna-Karin Brodin
Anna-Karin Pernestig
Gunnar Jacobsson
Rune Andersson
Diana Karlsson
Source
BMC Infect Dis. 2015;15:199
Date
2015
Language
English
Publication Type
Article
Keywords
Adult
Aged
Aged, 80 and over
Anti-Bacterial Agents - therapeutic use
DNA, Bacterial - analysis
Emergency Service, Hospital
Female
Fungi - isolation & purification
Gram-Negative Bacteria - isolation & purification
Gram-Positive Bacteria - isolation & purification
Humans
Male
Middle Aged
Multiplex Polymerase Chain Reaction
Nucleic Acid Amplification Techniques - standards
Patient Admission
Polymerase Chain Reaction - methods
Predictive value of tests
Reagent Kits, Diagnostic - standards
Sepsis - blood - diagnosis - drug therapy - microbiology
Sweden
Young Adult
Abstract
Sepsis is a serious medical condition requiring timely administered, appropriate antibiotic therapy. Blood culture is regarded as the gold standard for aetiological diagnosis of sepsis, but it suffers from low sensitivity and long turnaround time. Thus, nucleic acid amplification tests (NAATs) have emerged to shorten the time to identification of causative microbes. The aim of the present study was to evaluate the clinical utility in everyday practice in the emergency department of two commercial NAATs in patients suspected with sepsis.
During a six-week period, blood samples were collected consecutively from all adult patients admitted to the general emergency department for suspicion of a community-onset sepsis and treated with intravenous antibiotics. Along with conventional blood cultures, multiplex PCR (Magicplex™) was performed on whole blood specimens whereas portions from blood culture bottles were used for analysis by microarray-based assay (Prove-it™). The aetiological significance of identified organisms was determined by two infectious disease physicians based on clinical presentation and expected pathogenicity.
Among 382 episodes of suspected sepsis, clinically relevant microbes were detected by blood culture in 42 episodes (11%), by multiplex PCR in 37 episodes (9.7%), and by microarray in 32 episodes (8.4%). Although moderate agreement with blood culture (kappa 0.50), the multiplex PCR added diagnostic value by timely detection of 15 clinically relevant findings in blood culture-negative specimens. Results of the microarray corresponded very well to those of blood culture (kappa 0.90), but were available just marginally prior to blood culture results.
The use of NAATs on whole blood specimens in adjunct to current culture-based methods provides a clinical add-on value by allowing for detection of organisms missed by blood culture. However, the aetiological significance of findings detected by NAATs should be interpreted with caution as the high analytical sensitivity may add findings that do not necessarily corroborate with the clinical diagnosis.
Notes
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PubMed ID
25928122 View in PubMed
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Comparison of three multiplex real-time PCR assays for detection of enteric viruses in patients with diarrhea.

https://arctichealth.org/en/permalink/ahliterature299849
Source
Eur J Clin Microbiol Infect Dis. 2019 Feb; 38(2):241-244
Publication Type
Comparative Study
Journal Article
Date
Feb-2019
Author
Jari J Hirvonen
Author Affiliation
Fimlab Laboratories, Arvo Ylpön katu 4, FI-33520, Tampere, Finland. hirvonen.j.jari@gmail.com.
Source
Eur J Clin Microbiol Infect Dis. 2019 Feb; 38(2):241-244
Date
Feb-2019
Language
English
Publication Type
Comparative Study
Journal Article
Keywords
Adolescent
Adult
Aged
Aged, 80 and over
Child
Child, Preschool
Diarrhea - diagnosis
Feces - virology
Finland
Gastroenteritis - diagnosis
Humans
Middle Aged
Molecular Diagnostic Techniques - methods
Multiplex Polymerase Chain Reaction - methods
Prospective Studies
Reagent kits, diagnostic
Time Factors
Viruses - isolation & purification
Young Adult
Abstract
In this study, the usability and performance of three commercially available multiplex real-time RT-PCR assays for the detection of major enteric viruses were investigated. In total, 481 fecal specimens were analyzed using the Allplex™ GI Virus Assay, the Rida®Gene Viral Stool Panel I, and the FTD Viral Gastroenteritis. The overall agreement between the assays was 99.9%. Despite convergent analytical performance, differences between the multiplex RT-PCR assays were apparent when considering their suitability for routine diagnostics.
PubMed ID
30414091 View in PubMed
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Detection of PMS2 Mutations by Screening Hereditary Nonpolyposis Colon Cancer Families from Denmark and Sweden.

https://arctichealth.org/en/permalink/ahliterature310019
Source
Genet Test Mol Biomarkers. 2019 Sep; 23(9):688-695
Publication Type
Journal Article
Date
Sep-2019
Author
Henrik Okkels
Kristina Lagerstedt-Robinsson
Friedrik P Wikman
Thomas V O Hansen
Ihab Lolas
Lars Joachim Lindberg
Henrik B Krarup
Author Affiliation
Section of Molecular Diagnostics, Department of Clinical Chemistry, Aalborg University Hospital, Aalborg, Denmark.
Source
Genet Test Mol Biomarkers. 2019 Sep; 23(9):688-695
Date
Sep-2019
Language
English
Publication Type
Journal Article
Keywords
Animals
COS Cells
Chlorocebus aethiops
Colorectal Neoplasms - diagnosis - genetics
Colorectal Neoplasms, Hereditary Nonpolyposis - diagnosis - genetics
DNA Mutational Analysis
Denmark
Early Detection of Cancer
High-Throughput Nucleotide Sequencing
Mismatch Repair Endonuclease PMS2 - genetics
Multiplex Polymerase Chain Reaction
Polymerase Chain Reaction
Sweden
Abstract
Background and Aims: Hereditary nonpolyposis colon cancer (HNPCC) and Lynch syndrome (LS) are characterized by defects in the mismatch repair (MMR) system, which protects the integrity of the genome. Pathogenic variants in four MMR genes (MLH1, MSH2, MSH6, and PMS2) are responsible for LS, an autosomal, dominant hereditary disease that occurs with a frequency of 2-5% among all colorectal cancer cases. It has been estimated that ~2-5% of all pathogenic variants found in the four MMR genes in LS cases are detected in the PMS2 gene. An overview of detected variants is presented here. Materials and Methods: Long-range (LR) PMS2 polymerase chain reaction (PCR) and PMS2 multiplex ligation probe amplification (MLPA) assays were used to detect PMS2 variants in ~1500 probands. In a subset of the probands, pathogenic PMS2 variants were detected by next-generation sequencing, and all detected variants were confirmed by LR-PCR combined with an MLPA assay. Results: A summary of PMS2 mutation analyses performed on colon cancer patients from molecular diagnostic laboratories in Denmark and Sweden is presented. By screening ~1500 HNPCC probands, a total of 40 different PMS2 variants were detected in 71 probands (5%); 20 variants were classified as pathogenic (C5), 2 variants as likely pathogenic (C4), 15 variants as variants of unknown significance (VUSs) (C3), 1 variant as likely benign (C2), and 2 variants as benign (C1). In total, 22/71 (31%) of the probands carried a pathogenic sequence variant. Among the probands with isolated loss of pPMS2 expression, the fraction of pathogenic variants was 20/35 (55%). Conclusions: Approximately 5% of the probands found in the Danish and Swedish populations presented here carried a PMS2 variant. In this study, six novel pathogenic variants and seven VUSs are reported.
PubMed ID
31433215 View in PubMed
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Development of a multiplex PCR for identification of Dictyocaulus lungworms in domestic and wild ruminants.

https://arctichealth.org/en/permalink/ahliterature272741
Source
Parasitol Res. 2015 Oct;114(10):3923-6
Publication Type
Article
Date
Oct-2015
Author
Anna M Pyziel
Zdzislaw Laskowski
Johan Höglund
Source
Parasitol Res. 2015 Oct;114(10):3923-6
Date
Oct-2015
Language
English
Publication Type
Article
Keywords
Animals
Animals, Domestic - parasitology
Animals, Wild - parasitology
Cattle - parasitology
Deer - parasitology
Dictyocaulus - classification - genetics - isolation & purification
Dictyocaulus Infections - parasitology
Multiplex Polymerase Chain Reaction - methods
Poland
Ruminants - parasitology
Species Specificity
Sweden
Abstract
Dictyocaulus lungworms are the causative agents of parasitic bronchitis (dictyocaulosis) characterised by coughing and severe lung pathology in domestic and wild ruminants. The objective of this study was to design a simple molecular test that could detect of lungworm DNA from both adult and larval lungworms and could distinguish between the most common Dictyocaulus species found in cattle and in some species of wild ruminants. A multiplex PCR test with four novel primers targeting species-specific regions of the second internal transcribed spacer (ITS2) was designed based on our own sequence data as well as on available sequence information in GenBank. After PCR amplification of lungworms from European bison (Bison bonasus), cattle (Bos taurus), moose (Alces alces), red deer (Cervus elaphus) and roe deer (Capreolus capreolus), products were analysed with gel electrophoresis. This resulted in three specific bands of different size depending on the species analysed. Dictyocaulus viviparus collected from cattle or European bison resulted in a ca. 560 bp band, D. capreolus collected from roe deer produced a band ca. 400 bp and the longest DNA band (ca. 660 bp) was obtained with DNA from Dictyocaulus sp. collected from red deer and moose. Dictyocaulus eckerti bands with expected size of 714 bp were not observed in our study. The multiplex method produced consistent results with samples from both Sweden and Poland and overcame the limitations of traditional techniques based on differences in morphological features of parasites at different life stages.
Notes
Erratum In: Parasitol Res. 2015 Oct;114(10):393926323579
PubMed ID
26266883 View in PubMed
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Dominant optic atrophy in Denmark - report of 15 novel mutations in OPA1, using a strategy with a detection rate of 90%.

https://arctichealth.org/en/permalink/ahliterature121993
Source
BMC Med Genet. 2012;13:65
Publication Type
Article
Date
2012
Author
Gitte J Almind
Jakob Ek
Thomas Rosenberg
Hans Eiberg
Michael Larsen
Lucamp Lucamp
Karen Brøndum-Nielsen
Karen Grønskov
Author Affiliation
Center for Applied Human Molecular Genetics, Kennedy Center, Glostrup, Denmark. gij@kennedy.dk
Source
BMC Med Genet. 2012;13:65
Date
2012
Language
English
Publication Type
Article
Keywords
Base Sequence
Cohort Studies
DNA Primers - genetics
Denmark - epidemiology
Founder Effect
GTP Phosphohydrolases - genetics
Genetic Testing
Haplotypes - genetics
Humans
Molecular Sequence Data
Multiplex Polymerase Chain Reaction
Mutation - genetics
Optic Atrophy, Autosomal Dominant - epidemiology - genetics
Polymerase Chain Reaction
Polymorphism, Single Nucleotide - genetics
Sequence Analysis, DNA
Abstract
Investigation of the OPA1 mutation spectrum in autosomal dominant optic atrophy (ADOA) in Denmark.
Index patients from 93 unrelated ADOA families were assessed for a common Danish founder mutation (c.2826_2836delinsGGATGCTCCA) inOPA1. If negative, direct DNA sequencing of the coding sequence and multiplex ligation-dependent probe amplification (MLPA) were performed. Results from MLPA analysis have been previously reported. Haplotype analysis was carried out analysing single nucleotide polymorphisms (SNP). Retrospective clinical data were retrieved from medical files.
Probably causative mutations were identified in 84 out of 93 families (90%) including 15 novel mutations. Three mutations c.983A?>?G, c.2708_2711delTTAG and c.2826_2836delinsGGATGCTCCA, were responsible for ADOA in10, 11 and 28 families, respectively, corresponding to 11%, 12% and 30%. A common haplotype in nine of ten c.983A?>?G families suggests that they descend from a single founder. The c.2708_2711delTTAG mutation was present on at least two haplotypes and has been repeatedly reported in various ethnic groups,thus represents a mutational hotspot. Clinical examinations of index patients with the two latter mutations demonstrated large inter- and intra-familial variations apparently.
Genetic testing for OPA1mutations assist in the diagnosis. We have identified mutations in OPA1 in 90% of families including 15 novel mutations. Both DNA sequencing and MLPA analysis are necessary to achieve a high detection rate. More than half of the affected families in Denmark are represented by three common mutations, at least two of which are due to a founder effect, which may account for the high prevalence of ADOA in Denmark.
Notes
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PubMed ID
22857269 View in PubMed
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[Experience of application of multiplex qPCR for differential diagnostics of intestinal viral infections].

https://arctichealth.org/en/permalink/ahliterature117437
Source
Zh Mikrobiol Epidemiol Immunobiol. 2012 Nov-Dec;(6):39-45
Publication Type
Article
Author
A A Marova
A S Oksanich
A N Kaira
E R Meskina
E A Medvedeva
O E Ivanova
A N Lukashev
K K Kyuregian
M A Kalinkina
O V Egorova
V V Zverev
E V Faizuloev
Source
Zh Mikrobiol Epidemiol Immunobiol. 2012 Nov-Dec;(6):39-45
Language
Russian
Publication Type
Article
Keywords
Adenoviridae - genetics - isolation & purification
Adult
Child
Diagnosis, Differential
Enterovirus - genetics - isolation & purification
Female
Fluorescence
Hepatovirus - genetics - isolation & purification
Humans
Intestines - virology
Male
Mamastrovirus - genetics - isolation & purification
Multiplex Polymerase Chain Reaction - methods
Norovirus - genetics - isolation & purification
Orthoreovirus - genetics - isolation & purification
Poliovirus - genetics - isolation & purification
Reagent kits, diagnostic
Real-Time Polymerase Chain Reaction - methods
Rotavirus - genetics - isolation & purification
Russia - epidemiology
Sapovirus - genetics - isolation & purification
Virus Diseases - diagnosis - epidemiology - virology
Abstract
Evaluate the effectiveness of multiplex reverse transcription (RT) and polymerase chain reaction with fluorescence detection in real time mode (qPCR) methods for differential detection of 11 groups of intestine viruses (adenoviruses, enteroviruses, polioviruses, hepatitis A and E viruses, group A and C rotaviruses, orthoreoviruses, noroviruses, sapoviruses and astroviruses) in various biological samples.
Panels of virus isolates and clinical samples characterized by reference methods were used to evaluate sensitivity of detection of various intestine viruses. Nucleic acids were isolated from study samples and multiplex RT and qPCR were carried out.
Sensitivity of laboratory reagent kit (LRK) when compared with results obtained from reference methods was 100% for rotavirus A, adenovirus, enterovirus and norovirus, 88.9% for hepatitis E virus and 92.3% for hepatitis A virus, and diagnostic specificity - 99.4%. During analysis of 697 clinical samples from patients with acute intestine infection symptoms nucleic acids of various intestine viruses were isolated in 71.7%.
Multiplex qRT-PCR was shown as an effective method of etiologic diagnostics of an intestine viral infection. Use of LRK was demonstrated to establish etiology of intestine diseases in 63 - 72% and in children with watery diarrhea - in approximately 90% of cases.
PubMed ID
23297630 View in PubMed
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