Molecular characterization of hospital-acquired adenovirus infantile respiratory infection in Chile using species-specific PCR assays

2007 ◽  
Vol 39 (3) ◽  
pp. 175-181 ◽  
Author(s):  
Carmen Larrañaga ◽  
Jorge Martínez H. ◽  
Angélica Palomino M. ◽  
Mónica Peña C. ◽  
Flavio Carrión A. ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Respiratory Infection ◽  
Specific Pcr ◽  
Pcr Assays ◽  
Species Specific ◽  
Hospital Acquired

scholarly journals Molecular Characterization of Giardia duodenalis in Children and Adults Sampled in Algeria

2020 ◽  
Vol 9 (1) ◽  
pp. 54
Author(s):  
Salem Belkessa ◽  
Daniel Thomas-Lopez ◽  
Karim Houali ◽  
Farida Ghalmi ◽  
Christen Rune Stensvold
Keyword(s):  
Real Time ◽  
Molecular Characterization ◽  
Real Time Pcr ◽  
Intestinal Parasites ◽  
Giardia Duodenalis ◽  
Specific Pcr ◽  
Stool Samples ◽  
Pcr Targeting ◽  
Assemblage B

The molecular epidemiology of giardiasis in Africa remains unclear. A study was carried out across four hospitals in Algeria. A total of 119 fecal samples from 55 children, 37 adults, and 27 individuals of undetermined age, all scored positive for intestinal parasites by microscopy, and were screened by real-time PCR for Giardia. Molecular characterization of Giardia was performed by assemblage-specific PCR and PCR targeting the triose phosphate isomerase gene (tpi). Of the 119 samples, 80 (67%) were Giardia-positive by real-time PCR. For 48 moderately-highly real-time PCR-positive samples, tpi genotyping assigned 22 samples to Assemblage A and 26 to Assemblage B. Contrary to Assemblage A, Assemblage B exhibited substantial genetic diversity and allelic heterozygosity. Assemblage-specific PCR proved to be specific for discriminating Assemblage A or B but not as sensitive as tpi genotyping. We confirmed that real-time PCR is more sensitive than microscopy for detecting Giardia in stool samples and that robust amplification and sequencing of the tpi gene is feasible when moderate-to-strongly real-time PCR-positive samples are used. This study is one of the few performed in Africa providing genotyping data on Giardia infections in humans. Both assemblages A and B were commonly seen and not associated with specific sociodemographic data.

scholarly journals Development and Validation of a Nested-PCR-Denaturing Gradient Gel Electrophoresis Method for Taxonomic Characterization of Bifidobacterial Communities

2003 ◽  
Vol 69 (11) ◽  
pp. 6380-6385 ◽  
Author(s):  
R. Temmerman ◽  
L. Masco ◽  
T. Vanhoutte ◽  
G. Huys ◽  
J. Swings
Keyword(s):  
Nested Pcr ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Temporal Changes ◽  
Rrna Gene ◽  
Specific Pcr ◽  
Gradient Gel Electrophoresis ◽  
Species Specific ◽  
Taxonomic Characterization

ABSTRACT The taxonomic characterization of a bacterial community is difficult to combine with the monitoring of its temporal changes. None of the currently available identification techniques are able to visualize a “complete” community, whereas techniques designed for analyzing bacterial ecosystems generally display limited or labor-intensive identification potential. This paper describes the optimization and validation of a nested-PCR-denaturing gradient gel electrophoresis (DGGE) approach for the species-specific analysis of bifidobacterial communities from any ecosystem. The method comprises a Bifidobacterium-specific PCR step, followed by purification of the amplicons that serve as template DNA in a second PCR step that amplifies the V3 and V6-V8 regions of the 16S rRNA gene. A mix of both amplicons is analyzed on a DGGE gel, after which the band positions are compared with a previously constructed database of reference strains. The method was validated through the analysis of four artificial mixtures, mimicking the possible bifidobacterial microbiota of the human and chicken intestine, a rumen, and the environment, and of two fecal samples. Except for the species Bifidobacterium coryneforme and B. indicum, all currently known bifidobacteria originating from various ecosystems can be identified in a highly reproducible manner. Because no further cloning and sequencing of the DGGE bands is necessary, this nested-PCR-DGGE technique can be completed within a 24-h span, allowing the species-specific monitoring of temporal changes in the bifidobacterial community.

scholarly journals Technical note: Development of multiplex PCR assays for the molecular characterization of Streptococcus uberis strains isolated from bovine mastitis

2020 ◽  
Vol 103 (1) ◽  
pp. 915-921
Author(s):  
Dario Calonzi ◽  
Alicia Romano ◽  
Valentina Monistero ◽  
Paolo Moroni ◽  
Mario Vittorio Luini ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Molecular Characterization ◽  
Bovine Mastitis ◽  
Technical Note ◽  
Streptococcus Uberis ◽  
Pcr Assays

Molecular characterization of Schistosoma haematobium species-specific diagnostic antigen (gp23) using cDNA library in E. coli

2015 ◽  
Vol 27 (2) ◽  
pp. 299-310
Author(s):  
Emad A. Abada ◽  
Mohamed A. Al Abboud ◽  
Zeinat K. Mohamed ◽  
Maged M. Al-Sherbeiny
Keyword(s):  
Cdna Library ◽  
Molecular Characterization ◽  
Schistosoma Haematobium ◽  
E Coli ◽  
Diagnostic Antigen ◽  
Species Specific

scholarly journals Isolation and Characterization of Brachyspira spp. from Dogs in the Czech Republic

2010 ◽  
Vol 79 (3) ◽  
pp. 437-442
Author(s):  
Daniel Šperling ◽  
František Čada ◽  
Alois Čížek
Keyword(s):  
Czech Republic ◽  
Biochemical Activity ◽  
Animal Shelters ◽  
The Czech Republic ◽  
Specific Pcr ◽  
Isolation And Characterization ◽  
High Prevalence ◽  
Species Specific

The objectives of this study were to establish the prevalence of intestinal Spirochetes of the genusBrachyspirain Czech dogs and to determine the susceptibility of obtainedB. pilosicoliisolates to selected antibacterial substances. Spirochetes were diagnosed microscopically in 23 out of 1139 samples of dogs’ excrements, primarily intended for a parasitological testing. The cultivation of positive samples provided 10 brachyspira isolates, which were, on the basis of their biochemical activity and the results of the species-specific PCR, identified asB. pilosicoli(9 isolates) andB. hyodysenteriae(1 isolate). These dogs came from households. All the 7 tested isolatesB. pilosicoliwere sensitive to metronidazole and doxycycline, uniformly resistant to erythromycin, partly sensitive to cefazoline, lincomicine and ampicilline except for one isolate ofB. pilosicoli, which was resistant to ampicilline. The second part of study was focused on dogs with diarrhoea that came from animal shelters, where a high prevalence of 58% (10/17) ofB. pilosicoliwas found.

Validation of RT-PCR Assays for Molecular Characterization of Porcine Teschoviruses and Enteroviruses

2006 ◽  
Vol 53 (6) ◽  
pp. 257-265 ◽  
Author(s):  
G. La Rosa ◽  
M. Muscillo ◽  
A. Di Grazia ◽  
S. Fontana ◽  
M. Iaconelli ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Rt Pcr ◽  
Pcr Assays

scholarly journals Species-Specific PCR Assays for Differentiating Heterodera filipjevi and H. avenae

Plant Disease ◽  
2013 ◽  
Vol 97 (12) ◽  
pp. 1611-1619 ◽  
Author(s):  
Guiping Yan ◽  
Richard W. Smiley ◽  
Patricia A. Okubara ◽  
Andrea M. Skantar
Keyword(s):  
Pacific Northwest ◽  
Management Practices ◽  
Pcr Amplification ◽  
Nematode Species ◽  
Heterodera Avenae ◽  
Cyst Nematodes ◽  
Specific Pcr ◽  
The Pacific ◽  
Pcr Assays ◽  
Species Specific

Heterodera avenae and H. filipjevi are economically important cyst nematodes that restrict production of cereal crops in the Pacific Northwest United States and elsewhere in the world. Identification of these two species is critical for recommending and implementing effective management practices. Primers were designed from the internal transcribed spacer (ITS) regions of H. avenae and H. filipjevi ribosomal DNA. The primers were highly specific when examined on target isolates but did not amplify DNA from nontarget Heterodera, Globodera, Meloidogyne, Pratylenchus, and other nematode species tested. Polymerase chain reaction (PCR) and amplification conditions were established, and H. avenae and H. filipjevi were clearly distinguished by PCR fragments of 242 and 170 bp, respectively. Robust PCR amplification was achieved with DNA extracted from a single egg or second-stage juvenile (J2) using a laboratory-made worm lysis buffer, and DNA from 0.5 egg or J2 using a commercial kit. The PCR assays were successfully employed for differentiation of H. filipjevi and H. avenae populations collected from eight locations in three Pacific Northwest states. This is the first report of a species-specific ITS PCR assay to detect and identify H. filipjevi. The assays for both species will enhance diagnosis of cereal cyst nematode species in infested fields.

PCR SURVEILLANCE OF FOUR PROTOTYPIC RAT PARVOVIRUSES IN LABORATORY RAT COLONIES IN TAIWAN

2020 ◽  
Vol 46 (01) ◽  
pp. 29-35 ◽  
Author(s):  
Yung-Hui Chang ◽  
Yi-Chun Liao ◽  
Ko-Wei Wang ◽  
Ming-Hseng Wang ◽  
Ber-Hsiung Fang ◽  
...  
Keyword(s):  
Monitoring Program ◽  
Dual Infection ◽  
Virus Species ◽  
Laboratory Rats ◽  
Specific Pcr ◽  
Laboratory Rat ◽  
Minute Virus ◽  
Pcr Assays ◽  
Species Specific ◽  
Infectious Condition

Prototypic rat parvoviruses are among the most common infectious agents in laboratory rats worldwide. It is important to determine the infectious condition of rat parvoviruses in research colonies in Taiwan. In this study, virus species-specific PCR assays were applied to screen Kilham rat virus (KRV), Toolan’s H-1 virus (H-1 virus), rat parvovirus (RPV) and rat minute virus (RMV) in 203 rats from five different research colonies in Taiwan. In this survey, the infection rate of rat parvoviruses was 19.2% (39/203), including KRV (12.3%; 25/203), RMV (8.9%; 18/203) and RPV (1.0%; 2/203). Dual infection of KRV and RMV were detected in 3.0% (6/203) of samples. No rats were positive for H-1 virus. The infectious rates of KRV and RMV detected in this study are much higher than those reported in USA and in Europe. Rat parvovirus PCR assays reported in this paper could be a useful tool for routine rat health monitoring program applied in laboratory rat facilities. [Chang YH, Liao YC, Wang KW, Hsieh SC, Wang MH, Fang BH, Chang CY, Chueh LL, Wan CH, PCR Surveillance of four prototypic rat parvoviruses in laboratory rat colonies in Taiwan, Taiwan Vet J 46(1): 1–7, 2020]

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