• 제목/요약/키워드: Conventional Polymerase Chain Reaction

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Conservative treatment of cytomegalovirus colitis with bowel perforation in an immunocompetent patient: case report and review of literature

  • Nam, Kyoung Sik;Park, Hee Ug;Park, Min Gi;Park, Su Ho;Hwang, Ji Yeon;Kim, Dong Kyu;Kim, Sung Jun
    • Journal of Yeungnam Medical Science
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    • 제34권1호
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    • pp.75-79
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    • 2017
  • Cytomegalovirus (CMV) colitis, which is rare in an immunocompetent patient, was encountered in a 67-year-old man who was admitted due to persistent diarrhea. The first diagnostic tool was colonoscopy, which showed multiple ulcers from cecum to rectum. The secondary tool was CMV polymerase chain reaction, and CMV colitis was diagnosed. Intravenous ganciclovir therapy was administered, which resulted in improvement of diarrhea and ulcers throughout the colon were healed. Asymptomatic colon perforation was detected during diagnostic testing, which improved over the conventional treatment. CMV colitis is rare in immunocompetent patients, but it is essential for the differential diagnosis.

광바이오센서를 이용한 우유 및 유제품의 식중독균 신속검출법 (Rapid Detection Methods for Food-Borne Pathogens in Milk and Dairy Products using an Optical Biosensor)

  • 최은영;장진희;홍성욱;김소영;배효주;박범영;오미화
    • Journal of Dairy Science and Biotechnology
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    • 제31권2호
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    • pp.165-170
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    • 2013
  • Milk and dairy products are not only excellent foods for humans, providing plentiful varied nutrients, but are also a good medium for detrimental food-borne pathogens. Although the food safety field has stabilized due to standardization of food processing, such as the hazard analysis critical control point (HACCP), outbreaks and cases caused by food-borne pathogens still occur at high rates. In approximately 30% of cases, the disease-causing pathogenic organism is undetermined. Recently, a biosensor was developed that has a simple and fast response and overcomes the problems of conventional methods such as cultivation, immuno-assay, polymerase chain reaction, and microarray. Due to the high selectivity and sensitivity of optical biosensors, it is a suitable method for the immediate detection of food-borne pathogens in milk and dairy products.

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Towards developing a diagnostic regimen for the treatment follow-up of Trypanosoma brucei gambiense

  • Mbati, Peter-A.;Hirumi, Kazuko;Inoue, Noboru;Situakibanza, Nanituma-H.;Hirumi, Hiroyuki
    • Parasites, Hosts and Diseases
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    • 제37권4호
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    • pp.289-292
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    • 1999
  • BALB/c mice infected with a high virulent strain of Trypanosoma brucei gambiense IL3707 were treated intraperitoneally (ip) with either Melarsoprol (Mel-B) or PSG(+) buffer as controls. The mice were subsequently monitored regularly for parasites by direct microscopic examination of their tail blood or buffy coat and by polymerase chain reaction (PCR). Mel-B was found to be an effective drug for treatment against T.b. gambiense because at the end of the first treatment schedule, all treated mice were negative for parasites even by PCR, while all the control animals were positive. Three of the five Mel-B treated mice, while parasitologically negative, were PCR positive between 53 and 80 days post infection (DPI), indicating that they still harbored an infection. All treated mice were subsequently negative for parasites even by PCR at 88 DPI. A combination of conventional microscopic examination and PCR offers a good prediction of cure following treatment of trypanosomosis.

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Development of a multiplex PCR to identify Salmonella, Leptospira and Brucella species in tissue samples

  • Truong, Quang Lam;Yoon, Byung-Il;Hahn, Tae-Wook
    • 대한수의학회지
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    • 제52권2호
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    • pp.75-82
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    • 2012
  • We have developed and optimized a multiplex polymerase chain reaction (mPCR) for simultaneous detection of Brucella, Salmonella and Leptospira with high sensitivity and specificity. Three pairs of oligonucleotide primers were designed to specifically amplify the targeted genes of Salmonella, Leptospira and Brucella species with sizes of 521, 408 and 223 bp, respectively. The mPCR did not produce any nonspecific amplification products when tested against 15 related species of bacteria. The sensitivity of the mPCR was 100 fg for Brucella and 1 pg for both Salmonella and Leptospira species. In the field application, kidney, liver and spleen were collected from wild rats and stray cats and examined by mPCR. The high specificity and sensitivity of this mPCR assay provide a valuable tool for diagnosis and for the simultaneous and rapid detection of three zoonotic bacteria that cause disease in both humans and animals. Therefore, this assay could be a useful alternative to the conventional method of culture and single PCR for the detection of each pathogen.

Utility of taxon-specific molecular markers for the species identification of herbarium specimens: an example from Desmarestia japonica (Phaeophyceae, Desmarestiales) in Korea

  • Lee, Sang-Rae;Lee, Eun-Young
    • Fisheries and Aquatic Sciences
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    • 제21권3호
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    • pp.8.1-8.6
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    • 2018
  • Desmarestia japonica (Phaeophyceae, Desmarestiales) was recently established from the Japanese ligulate Desmarestia and is morphologically similar to D. ligulata. This species has been reported only from Japan. However, the taxonomic reports based on additional regional distributions are needed to clarify this taxonomic entity and its species boundaries. Because Desmarestia species have restricted distributions in Korea, we reexamined herbarium specimens of D. ligulata deposited at the National Institute of Biological Resources (South Korea). To improve the amplification efficiency of the polymerase chain reaction and avoid contamination by the DNA of other organisms, we developed taxon-specific molecular markers suitable for DNA barcoding of Desmarestia species. Nuclear ribosomal small subunit RNA (18S rDNA) and mitochondrial cytochrome c oxidase 1 (cox1) regions were selected as target DNA. As a result, both were successfully isolated from herbarium specimens of D. japonica acquired over 10 years. These molecular markers provide useful genetic information for herbarium specimens for which conventional molecular analysis is challenging.

Fast High-throughput Screening of the H1N1 Virus by Parallel Detection with Multi-channel Microchip Electrophoresis

  • Zhang, Peng;Park, Guenyoung;Kang, Seong Ho
    • Bulletin of the Korean Chemical Society
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    • 제35권4호
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    • pp.1082-1086
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    • 2014
  • A multi-channel microchip electrophoresis (MCME) method with parallel laser-induced fluorescence (LIF) detection was developed for rapid screening of H1N1 virus. The hemagglutinin (HA) and nucleocapsid protein (NP) gene of H1N1 virus were amplified using polymerase chain reaction (PCR). The amplified PCR products of the H1N1 virus DNA (HA, 116 bp and NP, 195 bp) were simultaneously detected within 25 s in three parallel channels using an expanded laser beam and a charge-coupled device camera. The parallel separations were demonstrated using a sieving gel matrix of 0.3% poly(ethylene oxide) ($M_r$ = 8,000,000) in $1{\times}$ TBE buffer (pH 8.4) with a programmed step electric field strength (PSEFS). The method was ~20 times faster than conventional slab gel electrophoresis, without any loss of resolving power or reproducibility. The proposed MCME/PSEFS assay technique provides a simple and accurate method for fast high-throughput screening of infectious virus DNA molecules under 400 bp.

SHV ESBL생성 Klebsiella pneumoniae 균주의 실시간중합효소반응분석 (Real-Time PCR Analysis of SHV Extended-Spectrum beta-Lactamases Producing Klebsiella pneumoniae)

  • 양병선;육근돌
    • 대한임상검사과학회지
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    • 제41권4호
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    • pp.153-157
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    • 2009
  • The production of extended-spectrum ${\beta}$-lactamases ($ESBL_S$) of the TEM or SHV type by bacterial pathogens is a major threat to the use of the clinically important expanded-spectrum cephalosporins. The characterization of the SHV ESBLs producing Klebsiella pneumoniae strains present in clinical isolates is time-consuming processes. We describe here in the development of a novel system, which consists of a real time PCR. We found 11 K. pneumoniae strains to be presumptive strains ESBLs producers by clinical and laboratory standards institute (CLSI) guidelines. The double disk synergy test showed 8 ESBL positive and conventional PCR showed 10 SHV ESBL positive, which were K. pneumoniae strains isolates. By real time PCR analysis, SHV gene in 11 of 11 strains were identified. When sequencing analysis was compared with real time PCR, both analysis were presented 99% similarity. In this study, we used a rapid, sensitive, and specific real-time PCR (RT-PCR) method for detection of the assay SHV ESBL producing K. pneumoniae strains in clinical isolates.

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Rapid PCR Method for Detecting Candida albicans Using Primers Derived from the Integrin-like Protein Gene $\alpha$INT1 of Candida albicans

  • Lim, Young-Hee;Lee, Do-Hyun
    • Journal of Microbiology
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    • 제38권2호
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    • pp.105-108
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    • 2000
  • Oligonucleotide primers amplifying a 344 bp fragment on the integrin-like protein alpha-INT1p gene (${\alpha}$INT1) of Candida albicans were synthesized for screenign of C. albicans from clinicalsamples by the polymerase chain reaction (PCR). The PCR specifically amplified DNA from C. albicans and none from any other Candida, fungal, or human DNA in standard used here. The PCR assay showed that the primers (LH1 and LH2) were specific for 26 isolates of C. albicans from clinical smaples, whereas the positive fragment, 344 bp, was not amplified from 15 clinical isolates including 14 other medically important Candida species and an isolate of Saccharomyces cerevisiae. PCR was conducted on the urine samples of 20 patients and 4 samples were C. albicans positive. The detection limit of the PCR assay for C. albicans was shown to be approximately 10 cells/ml saline. The PCR system using 344 bp ${\alpha}$INT1 as a target is more specific and rapid than the conventional culture method, and the sensitive detection method is applicable to clinical diagnosis of C. albicans infections.

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Development of cellulose nano beads based a rapid detection kit to detect staphylococcal enterotoxin B

  • Kim, Giyoung;Yoo, Jinyoung;Park, Saetbyeol
    • 농업과학연구
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    • 제46권3호
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    • pp.549-557
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    • 2019
  • Staphylococcal enterotoxin is a very common cause of food poisoning. Conventional detection methods for the toxin including enzyme-linked immunosorbent assays (ELISAs), chemiluminescence (ECL), and polymerase chain reaction (PCR) assays require a lot of time, efforts, and expert technicians. Lateral flow strip kits have shown great potential for the rapid detection of foodborne pathogens. The lateral flow strip kit is widely used in clinical settings because it is easy to use, fast, and cost effective. A typical lateral flow strip kit uses colloidal gold to generate a visual signal. However, the lateral flow strip kit based on colloidal gold has limited sensitivity to fulfill food safety regulation requirements. This study was performed to develop a rapid test kit for pathogenic staphylococcal enterotoxin B (SEB) in food samples. The rapid detection kit was fabricated based on a nitrocellulose lateral-flow strip. Cellulose nano beads and SEB antibodies were used as the tag and receptor, respectively, to improve the detection performance. Manually spotted SEB antibody and anti-rabbit antibody on the surface of the nitrocellulose membrane were used as test and control spots, respectively. The feasibility of the rapid test kit to detect SEB in samples was evaluated. The sensitivity of the kit was 10 ng/mL SEB spiked in PBS. Additionally, the rapid test kit could detect 1 ng/mL of SEB in chicken meat extract.

Ginsenoside Rg1 promotes browning by inducing UCP1 expression and mitochondrial activity in 3T3-L1 and subcutaneous white adipocytes

  • Lee, Kippeum;Seo, Young-Jin;Song, Ji-Hyoen;Chei, Sungwoo;Lee, Boo-Yong
    • Journal of Ginseng Research
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    • 제43권4호
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    • pp.589-599
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    • 2019
  • Background: Panax ginseng Meyer is known as a conventional herbal medicine, and ginsenoside Rg1, a steroid glycoside, is one of its components. Although Rg1 has been proved to have an antiobesity effect, the mechanism of this effect and whether it involves adipose browning have not been elucidated. Methods: 3T3-L1 and subcutaneous white adipocytes from mice were used to access the thermogenic effect of Rg1. Adipose mitochondria and uncoupling protein 1 (UCP1) expression were analyzed by immunofluorescence. Protein level and mRNA of UCP1 were also evaluated by Western blotting and realtime polymerase chain reaction, respectively. Results: Rg1 dramatically enhanced expression of brown adipocyte-especific markers, such as UCP1 and fatty acid oxidation genes, including carnitine palmitoyltransferase 1. In addition, it modulated lipid metabolism, activated 5' adenosine monophosphate (AMP)-activated protein kinase, and promoted lipid droplet dispersion. Conclusions: Rg1 increases UCP1 expression and mitochondrial biogenesis in 3T3-L1 and subcutaneous white adipose cells isolated from C57BL/6 mice. We suggest that Rg1 exerts its antiobesity effects by promoting adipocyte browning through activation of the AMP-activated protein kinase pathway.