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#28334789   2017/03/23 Save this To Up

Comparative Evaluation of Norovirus Infection in Children with Acute Gastroenteritis by Rapid Immunochromatographic Test, RT-PCR and Real-time RT-PCR.

Immunochromatographic (IC) test for norovirus detection is a rapid and simple detection method. This study evaluated the sensitivity and specificity of a recent version of R-Biopharm RIDA®QUICK Norovirus IC assay for norovirus detection in fecal specimens from children hospitalized with acute gastroenteritis. Fecal specimens were tested by IC kit in comparison with gold standard reverse transcription polymerase chain reaction (RT-PCR) and real-time RT-PCR. The IC kit showed high sensitivity and specificity comparable with PCR-based methods. None of false positive and false negative was found and the assay did not cross-react with other gastroenteritis viruses. The IC assay could detect genogroup I.5 (GI.5) and a wide range of genotypes in the GII noroviruses including GII.3, GII.4, GII.6, GII.7, GII.14, GII.15, GII.21, and also newly emerging GII.17 norovirus. In conclusion, this norovirus IC kit could be an alternative choice for rapid screening or a quick diagnostic tool for norovirus detection in fecal specimens of acute gastroenteritis patients.

1492 related Products with: Comparative Evaluation of Norovirus Infection in Children with Acute Gastroenteritis by Rapid Immunochromatographic Test, RT-PCR and Real-time RT-PCR.

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#27628591   2016/09/15 Save this To Up

Establishment of an on-site diagnostic procedure for detection of orf virus from oral lesions of Japanese serows (Capricornis crispus) by loop-mediated isothermal amplification.

Orf virus infection has been prevalent continuously in the population of wild Japanese serows (Capricornis crispus), goat-like grazing cloven-hoofed mammal species that live mainly in mountainous areas of Japan. Currently, definitive diagnosis of infection requires time-consuming laboratory work. To diagnose rapidly on-site, we developed a field-friendly procedure for the detection of orf virus from oral cavity lesions. DNA was extracted from goat saliva spiked with orf virus as a proxy for Japanese serows by a commercial kit without the use of electricity, and the quality of the extracted DNA was evaluated by conventional polymerase chain reaction (PCR). Extracted DNA was amenable to DNA amplification, the same as when extracted in a laboratory. Next, to find optimal conditions for DNA amplification by loop-mediated isothermal amplification (LAMP), Bst and Csa DNA polymerases and 3 colorimetric indicators for visual diagnosis, hydroxy naphthol blue (HNB), malachite green and D-QUICK, were compared using a portable cordless incubator. The combination of Bst or Csa DNA polymerase with HNB was found to be easiest for visual diagnosis by the naked eye, and viral DNA was successfully amplified from all orf virus strains used. These results suggest that the procedure established here can work completely on-site and can be useful for definitive diagnosis and differentiation of orf virus infection in Japanese serows in remote mountainous areas.

2804 related Products with: Establishment of an on-site diagnostic procedure for detection of orf virus from oral lesions of Japanese serows (Capricornis crispus) by loop-mediated isothermal amplification.

Ofloxacin CAS Number [824 MOUSE ANTI CANINE DISTEMP HEV (Birma) ORF2 recombin HEV (Birma) ORF2 recombin HEV (Birma) ORF2 recombin VZV ORF9 recombinant anti VZV ORF26 recombinant ant Recombinant Viral Antige Tick born encephalitis vi Rubella virus E1 mosaic r Rubella virus E2 recombin Rubella virus capsid (C)

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#27609584   2016/09/09 Save this To Up

[Construction of anti-B7-H4-scFv library and screening and identification of anti-B7-H4-scFv].

Objective To construct the ribosome display library of anti-B7-H4 extracellular domain, and select the antibody with high specificity. Methods The cDNA of B7-H4 extracellular domain was amplified from A549 cells by reverse transcription PCR (RT-PCR). To express ectodomains of B7-H4, the sequence of B7-H4 gene, which encodes the B7-H4 extracellular domains, was inserted into plasmid pET-28a(+). The purified recombinant protein of B7-H4 extracellular domain was used to immunize BALB/c mice. The total RNA was extracted from the spleen of BALB/c mice which had been immunized with B7-H4 recombinant protein. The genes of VH, Vκ and VH/Vκ were amplified separately by RT-PCR and splicing by overlap extension PCR (SOE-PCR). The gene of VH/ Vκ was ligated into pUM19-T vector and the ligated sample was transformed into competent E.coli DH5α. The resulting plasmid was isolated and then subjected to sequencing to verify the gene sequence. TNT(R)T7 Quick for PCR DNA kit was used to translate and screen the anti-B7-H4-scFv in vitro from the ribosome display library. Western blotting and an indirect ELISA were performed to detect the specificity of anti-B7-H4-scFv. Results The right sequences of VH, Vκ and VH/Vκ were acquired, which were 439, 680 and 1098 bp in length, respectively. The analysis of specificity demonstrated that the anti-B7-H4-scFv screened from the ribosome display library had a high specific combining ability with B7-H4. Conclusion The experiment has successively constructed the ribosome display library of anti-B7-H4 extracellular domain, and selected the anti-B7-H4-scFv which has a high specific binding ability with recombinant protein of B7-H4 extracelluar domain.

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#27451970   2016/07/25 Save this To Up

Investigation of Rotavirus with Various Methods in Children with Acute Gastroenteritis and Determination of Its Molecular Epidemiology in Kayseri Province, Turkey.

In this study, the fresh stool samples from 254 children under 5 years of age with acute gastroenteritis which were delivered between October 2012 and December 2013 were collected.

1830 related Products with: Investigation of Rotavirus with Various Methods in Children with Acute Gastroenteritis and Determination of Its Molecular Epidemiology in Kayseri Province, Turkey.

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#27350908   2016/06/28 Save this To Up

Detection of dengue, west Nile virus, rickettsiosis and leptospirosis by a new real-time PCR strategy.

Dengue virus (DENV) infection causes sudden fever along with several nonspecific signs and symptoms and in severe cases, death. DENV is transmitted to people by Aedes aegypti and Ae. albopictus mosquitoes, whose populations increase during rainy season. West Nile Virus (WNV), Rickettsia spp. and Leptospira spp. are fever-causing pathogens that share many of the initial symptoms of DENV infection and also thrive in the rainy season. Outbreaks in some regions may be due to any of these pathogens that can co-circulate. Plus, they are clinically indistinguishable until severe symptoms appear, even though these diseases should be treated differently. An effective differential diagnosis would help clinicians and vector control departments to make right decisions for control and treatment of these diseases. Therefore, we developed four different SYBR green (®) -based reverse transcription quantitative PCR (RT-qPCR) assays for simultaneous detection of DENV, WNV, Rickettsia spp. and Leptospira spp. The assay has been optimized to yield results in less than 1 h; and in order to reduce contamination risk, all reagents were premixed and lyophilized on 96 well plates and thus only requires the addition of water and total nucleic acids from the sample. Sensitivities of the assays were less than 100 copies of nucleic acid targeted for these four pathogens. Assays did not show cross reactivity with any of the four pathogens nor to human nucleic acids. We are presenting a sensitive and selective kit that detects four relevant pathogens from tropical regions, that is quick, cost-effective and easy to use.

1643 related Products with: Detection of dengue, west Nile virus, rickettsiosis and leptospirosis by a new real-time PCR strategy.

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#27174027   2016/08/11 Save this To Up

Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for equipment-free detection of Cryptosporidium spp. oocysts in dairy cattle feces.

Cryptosporidium is a widespread protozoan parasite that infects a large number of vertebrate animals, resulting in varying degrees of diarrhea or even death. As dairy cattle feces is an important source of Cryptosporidium spp. infection, development of a handy and accurate detection method via its oocysts in dairy cattle feces would be interesting and necessary. We herein developed a quick detecting method using recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip to detect DNA of Cryptosporidium oocysts in dairy cattle feces. The DNA was released by boiled water with 0.1 % N-lauroylsarcosine sodium salt (LSS). The established method was proven to be of higher sensitivity than normal polymerase chain reaction (PCR) amplification with the lowest detection of 0.5 oocyst per reaction, and specificity with no cross reactivity to other common protozoan species in the intestine of dairy cattle. The diagnostic method established herein is simple, rapid, and cost-effective, and has potential for further development as a diagnostic kit for the diagnosis of cryptosporidiosis of dairy cattle.

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#26806228   2016/04/11 Save this To Up

Optimising the diagnostic strategy for onychomycosis from sample collection to FUNGAL identification evaluation of a diagnostic kit for real-time PCR.

Onychomycosis is a common nail disorder mainly due to dermatophytes for which the conventional diagnosis requires direct microscopic observation and culture of a biological sample. Nevertheless, antifungal treatments are commonly prescribed without a mycological examination having been performed, partly because of the slow growth of dermatophytes. Therefore, molecular biology has been applied to this pathology, to support a quick and accurate distinction between onychomycosis and other nail damage. Commercial kits are now available from several companies for improving traditional microbiological diagnosis. In this paper, we present the first evaluation of the real-time PCR kit marketed by Bio Evolution for the diagnosis of dermatophytosis. Secondly, we compare the efficacy of the kit on optimal and non-optimal samples. This study was conducted on 180 nails samples, processed by conventional methods and retrospectively analysed using this kit. According to our results, this molecular kit has shown high specificity and sensitivity in detecting dermatophytes, regardless of sample quality. On the other hand, and as expected, optimal samples allowed the identification of a higher number of dermatophytes by conventional mycological diagnosis, compared to non-optimal samples. Finally, we have suggested several strategies for the practical use of such a kit in a medical laboratory for quick pathogen detection.

1582 related Products with: Optimising the diagnostic strategy for onychomycosis from sample collection to FUNGAL identification evaluation of a diagnostic kit for real-time PCR.

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#26506733   2015/10/28 Save this To Up

LABORATORY DIAGNOSIS OF DENGUE VIRUS INFECTIONS.

Dengue diagnosis was one of the topics discussed at "the adult dengue" presentations. In this paper, a review is presented focusing on the main challenges of dengue laboratory diagnosis. Accurate and efficient diagnosis of dengue is important for clinical care, surveillance support, pathogenesis studies, and vaccine research. Laboratory diagnosis is also important for case confirmation. Laboratory dengue diagnosis can be performed through virus isolation, genome and antigen detection and serological studies. For virus detection, dengue viremia is short, usually observed two or three days before onset of fever and lasts four to five days later. Therefore, samples for virus detection must be taken in the first four to five days of the disease during febrile phase. In recent years, PCR (polymerase chain reaction) has become an important tool as a quick method for diagnosis of dengue, another is detection of NS1 antigen, using commercial ELISA kit. Serological studies, for primary infection, the dominant immunoglobulin isotype is IgM, anti-IgM may appear during febrile phase (50% of cases), the other half, it appears within 2-3 days of defervescence. Once detectable, IgM levels rise quickly and appears to peak about 2 weeks after the onset of symptoms, then they decline to undetectable level over 2-3 months. Anti-IgG appears shortly afterwards with very low level. The physiological definition of a primary infection is therefore characterized by a high molar fraction of anti-dengue IgM and low molar fraction of IgG. Secondary dengue infections are characterized by a rapid increase in IgG antibodies, anti-dengue IgM appears in most instances, the level are dramatically lower.

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#26248054   2015/08/31 Save this To Up

Evaluation of the updated RIDA®QUICK (Version N1402) immunochromatographic assay for the detection of norovirus in clinical specimens.

The sensitivity and specificity of the R-Biopharm RIDA(®)QUICK (N1402) immunochromatography assay for norovirus detection was examined using fecal material from Australian gastroenteritis incidents. The study involved the analysis of 3 groups of specimens; group 1 comprised 100 norovirus open reading frame (ORF) 1 RT-PCR positive specimens; group 2 comprised 100 ORF 1 RT-PCR norovirus negative specimens and group 3 comprised 12 specimens containing common gastroenteritis viruses other than norovirus. The RIDA(®)QUICK (N1402) assay detected both GI and GII norovirus and had an overall sensitivity of 87%. Genotype analysis of the capsid region of the genome (ORF 2) indicated the RIDA(®)QUICK (N1402) assay could detect a range of genotypes including GI.1, GI.2, GI.3, GI.4, GI.5, GII.3, GII.4 (including variants GII.4 (2009-like), GII.4 (2012), GII.4 (2012-like) and GII.4 (unknown)), GII.6, GII.13 and GII.21. The assay had good sensitivity for both GI and GII norovirus. The assay had a specificity of 97% and did not cross react with a number of common fecal viruses. However, one of eight rotavirus positive, norovirus negative specimens gave a positive result; rotavirus cannot be taken as the cause of such a false positive but cannot be excluded either. The kit was quick and easy to use and would be valuable in point-of-care testing.

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#26166499   2016/05/23 Save this To Up

An Improved Multiplex Real-Time SYBR Green PCR Assay for Analysis of 24 Target Genes from 16 Bacterial Species in Fecal DNA Samples from Patients with Foodborne Illnesses.

Here, we developed a new version of our original screening system (Rapid Foodborne Bacterial Screening 24; RFBS24), which can simultaneously detect 24 genes of foodborne pathogens in fecal DNA samples. This new version (RFBS24 ver. 5) detected all known stx2 subtypes, enterotoxigenic Escherichia coli (STh genotype), and Vibrio parahaemolyticus (trh2), which were not detected by the original RFBS24 assay. The detection limits of RFBS24 ver. 5 were approximately 5.6 × 10(-2)-5.6 × 10(-5) (ng DNA)/reaction, significantly lower (10- to 100-fold) than those of the original RFBS24 for the 22 target genes analyzed here. We also tested the new assay on fecal DNA samples from patients infected with Salmonella, Campylobacter, or enterohemorrhagic E. coli. The number of bacterial target genes detected by RFBS24 ver. 5 was greater than that detected by RFBS24. RFBS24 ver. 5 combined with an Ultra Clean Fecal DNA Isolation Kit showed adequate performance (sensitivity and specificity 89% and 100%, respectively, for Salmonella spp. and 100% and 83%, respectively, for Campylobacter jejuni) in terms of rapid detection of a causative pathogen during foodborne-illness outbreaks. Thus, RFBS24 ver. 5 is more useful than the previous assay system for detection of foodborne pathogens and offers quick simultaneous analysis of many targets and thus facilitates rapid dissemination of information to public health officials.

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