• Title/Summary/Keyword: Detection/Identification

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Sequence and Phylogenetic Analysis of Respiratory Syncytial Virus Isolated from Korea (국내에서 유행한 Respiratory Syncytial 바이러스의 염기서열 및 계통분석)

  • Kwon, Soon-Young;Choi, Young-Ju;Kim, So-Youn;Song, Ki-Joon;Lee, Yong-Ju;Choi, Jong-Ouck;Seong, In-Wha
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.9-22
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    • 1996
  • Respiratory Syncytial virus (RSV) is an important cause of acute lower respiratory tract infections in human, with infants and young children being particularly susceptible. In the temperate zones, sharp annual outbreaks of RSV occur during the colder months, in both the northern and the southern hemisphere. RSV is unusual in that it can repeatedly reinfect individuals throughout life and infect babies in the presence of maternal antibody. RSV isolates can be divided into two subgroups, A and B, on the basis of their reactions with monoclonal antibodies, and the two subgroups are also distinct at the nucleotide sequence level. The specific diagnosis of RSV infection was best made by isolation of virus in tissue culture, identification of viral antigen, or by specific serologic procedures. Recently, rapid detection of RSV and analysis of RSV strain variation became possible by development of methods of reverse transcription and polymerase chain reaction amplification. In this study, to determine the genetic diversity of RSV found in Korea, 173 bp and 164 bp spanning selected regions of the RSV F and SH genes were enzymatically amplified and sequenced, respectively. Eight for F gene and three for SH gene were detected in 66 nasopharyngeal swap samples tested. Two major antigenic subgroups, A and B were confirmed from Korean samples (seven for subgroup A and one for subgroup B). At the nucleotide level of the F gene region, Korean subgroup A strains showed 95-99% homologies compared to the prototype A2 strain of subgroup A and 93-100% homologies among Korean subgroup A themselves. For the SH gene region, Korean subgroup A strain showed 97.5% homology compared to the prototype A2 strain of subgroup A, and Korean subgroup B strain showed 97% homology compared to the prototype 18537 strain of subgroup B. Most of base changes were transition and occured in codon position 3, which resulted in amino acid conservation. Using the maximum parsimony method, phylogenetic analysis indicated that Korean RSV strains formed a group with other RSV strains isolated from the United States, Canada, the Great Britain and Australia.

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Isolation and identification of Cryptosporidium from various animals in Korea I. Prevalence of Cyuptosporidium in various animals (한국산 동물로부터 크립토스포리디움의 분리 및 동정 I . 각종 동물의 크립토스포리더움 감염상황)

  • Lee, Jae-Gu;Seo, Yeong-Seok;Park, Bae-Geun
    • Parasites, Hosts and Diseases
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    • v.29 no.2
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    • pp.139-148
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    • 1991
  • Cryptosporidium, a coccidian protozoa, commonly causes a self-limiting diarrheal illness in humans and animals. Fecal samples from various animals in Chonbuk district were observed using Sheather's aoatation technique, Kinyoun's modified acid-fast staining, and osmic acid pre-fixed Giemsa staining. The oocysts were detected in 74 cages(29.6%) out of 250 cages of mature mice, 26 (13.3%) out of 195 mature house rats, 75 (15.0%) out of 4-week-old 500 fowls, 98(19.9%) out of 6 to 8-month-old 500 pigs, and 111(22,2%) out of 2 to 5-year-old 500 dairy cattle, respectively. The degree of prevalence was slight in general, but actual prevalence was higher than infection rate because the detection rates were higher in repeated-preparation examinations in comparison to the first examination. Meanwhile, large and small types of oocysts were detected from mice, house rats, pigs, and cattle, and midium type from fowls.

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Analysis of Chromosome aberrations by fluorescence in situ hybridization using triple chromosome-specific probes in human lymphocyte exposed to radiation (3중 DNA probe를 이용한 FISH(fluorescence in situ hybridization) 기법으로 방사선에 의한 염색체 이상 분석)

  • Chung, Hai-Won;Kim, Su-Young;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.24 no.1
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    • pp.45-53
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    • 1999
  • Fluorescence in situ hybridization with chromosome-specific probe has been shown to be a valid and rapid method for detection of chromosome rearrangements induced by radiation. This method is useful for quantifying structural aberrations, expecially for stable ones, such as translocation and insertion, which are difficult to detect with conventional method in human lymphocyte. In order to apply FISH method for high dose biological dosimetry, chromosomal abberations by radiation at doses of 1, 3, 5, and 7Gy were analysed with whole chromosome-specific probes by human chromosome 1, 2 and 4 according to PAINT system. The frequencies of stable translocation per cell equivalent were 0.04, 0.33, 1.22, 2.62, and 5.58 for the lymphocyte exposed to 0, 1, 3, 5, and 7Gy, respectively, and those of dicentric were 0.00, 0.06, 0.52, 1.19 and 2.44, respectively. Significantly more translocation of t(Ab), a translocated chromosome with a piece of painted acentric matrial 'b' attached to unpainted piece containing centromere 'A', than reciprocal chromosome t(Ba) was observed. The frequencies of all type of chromosome rearrangements increased with dose. From above result, FISH seemed to be useful for radiation biodosimetry by which the frequencies of various types of stable aberrations in human lymphocyte can be observed more easily than by conventional method and so will improve our ability to perform meaningful biodosimetry.

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Screening for Lynch Syndrome in Young Colorectal Cancer Patients from Saudi Arabia Using Microsatellite Instability as the Initial Test

  • Alqahtani, Masood;Grieu, Fabienne;Carrello, Amerigo;Amanuel, Benhur;Mashour, Miral;Alattas, Rabab;Al-Saleh, Khalid;Alsheikh, Abdulmalik;Alqahtani, Sarah;Iacopetta, Barry
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1917-1923
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    • 2016
  • Background: Lynch Syndrome (LS) is a familial cancer condition caused by germline mutations in DNA mismatch repair genes. Individuals with LS have a greatly increased risk of developing colorectal cancer (CRC) and it is therefore important to identify mutation carriers so they can undergo regular surveillance. Tumor DNA from LS patients characteristically shows microsatellite instability (MSI). Our aim here was to screen young CRC patients for MSI as a first step in the identification of unrecognized cases of LS in the Saudi population. Materials and Methods: Archival tumor tissue was obtained from 284 CRC patients treated at 4 institutes in Dammam and Riyadh between 2006 and 2015 and aged less than 60 years at diagnosis. MSI screening was performed using the BAT-26 microsatellite marker and positive cases confirmed using the pentaplex MSI analysis system. Positive cases were screened for BRAF mutations to exclude sporadic CRC and were evaluated for loss of expression of 4 DNA mismatch repair proteins using immunohistochemistry. Results: MSI was found in 33/284 (11.6%) cases, of which only one showed a BRAF mutation. Saudi MSI cases showed similar instability in the BAT-26 and BAT-25 markers to Australian MSI cases, but significantly lower frequencies of instability in 3 other microsatellite markers. Conclusions: MSI screening of young Saudi CRC patients reveals that approximately 1 in 9 are candidates for LS. Patients with MSI are strongly recommended to undergo genetic counselling and germline mutation testing for LS. Other affected family members can then be identified and offered regular surveillance for early detection of LS-associated cancers.

Analytical Method Development and Monitoring of Residual Solvents in Dietary Supplements (건강기능식품 중 잔류용매 분석법 개발 및 모니터링)

  • Lee, Hwa-Mi;Shin, Ji-Eun;Jang, Young-Mi;Kim, Hee-Yun;Kim, Mee-Hye
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.390-397
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    • 2010
  • Residual solvents in foods are defined as organic volatile chemicals used or produced in manufacturing of extracts or additives, or functional foods. The solvents are not completely eliminated by practical manufacturing techniques and they also may become contaminated by solvents from packing, transportation or storage in warehouses. Because residual solvents have no nutritional value but may be hazardous to human health, there is a need to remove them from the final products or reduce their amounts to below acceptable levels. The purpose of this study was to develop and evaluate an analytical method for the screening of residual solvents in health functional foods. Furthermore, the aim of this study was to constitute a reasonable management system based on the current state of the market and case studies of foreign countries. Eleven volatile solvents such as MeOH, EtOH, trichloroethylene and hexane were separated depending on their column properties, temp. and time using Gas Chromatography (GC). After determining the GC conditions, a sample preparation method using HSS (Head Space Sampling) was developed. From the results, a method for analyzing residual solvents in health functional foods was developed considering matrix effect and interference from the sample obtained from the solution of solvents-free health functional foods spiked with 11 standards solutions. Validation test using the developed GC/HSS/MS (Mass Spectrometry) method was followed by tests for precision, accuracy, recovery, linearity and adequate sensitivity. Finally, examination of 104 samples grouped in suits was performed by the developed HSS/GC/MS for screening the solvents. The 11 solvents were isolated from health functional foods based on vapor pressure difference, and followed by separation within 15 minutes in a single run. The limt of detection (LOD), limit of quantification (LOQ), recovery and coefficient of variation (C.V.) of these compounds determined by the HSS/GC/MS were found to be 0.1 pg/mL, 0.1-125 pg/g, 51.0-104.6%, and less than 15%, respectively. Using the developed HSS/GC/MS method, residual solvent from 16 out of 104 health functional products were detected as a EtOH. This method therefore seems t o be a valuable extension ofanalytical method for the identification of residual solvents in health functional food.

Identification of Hanwoo (Korean Native Cattle) Beef in Restaurants using Real-time PCR (시중 음식점에서 판매되는 쇠고기의 유전자 분석을 이용한 한우육 감별)

  • Kim Jin-Man;Nam Yong-Suk;Choi Ji-Hun;Lee Mi-Ae;Jeong Jong-Yon;Kim Cheon-Jei
    • Food Science of Animal Resources
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    • v.25 no.2
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    • pp.203-209
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    • 2005
  • Real time-polymerase chain reaction (RT-PCR) is currently considered as the most sensitive method to detect low abundant DNAs in samples. Compared to conventional PCR, real-time PCR has a high reliability because of excluding false-positive results and can allow a simultaneous faster detection and quantification of target DNAs. This study was carried out to identify the Hanwoo (Korean native cattle) beef by genotyping after DNA extraction of commercial beef in 41 restaurants. Since Hanwoo, Holstein and imported cattle meat have different patterns in the MC1R gene associated with the coat colors of cattles (C-type, C/T-type or T-type), we could identify the genotype using real-time PCR The result of real-time PCR assay for beef samples in 41 restaurants which are asserted to sell Hanwoo beef only, showed that 29 of 41 samples were Hanwoo beef gene type (T-type) and 12 of 41 samples were Holstein or imported cattle gene type (C-type or C/T-type). Therefore, the proportion of Han-woo beef was $70.7\%$ and the proportion of Holstein or imported cattle meat was $29.3\%(C/T-type; 12.2\%,\;C-type; 17.1\%)$.

Genetic and Morphologic Identification of Spirometra ranarum in Myanmar

  • Jeon, Hyeong-Kyu;Park, Hansol;Lee, Dongmin;Choe, Seongjun;Kang, Yeseul;Bia, Mohammed Mebarek;Lee, Sang-Hwa;Sohn, Woon-Mok;Hong, Sung-Jong;Chai, Jong-Yil;Eom, Keeseon S.
    • Parasites, Hosts and Diseases
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    • v.56 no.3
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    • pp.275-280
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    • 2018
  • In the present study, we identified a Spirometra species of Myanmar origin (plerocercoid) by molecular analysis using mitochondrial cox1 and nad1 genes, as well as by morphological observations of an adult tapeworm. Spargana specimens were collected from a paddy-field in Taik Kyi Township Tarkwa Village, Yangon, Myanmar in December 2017. A total of 5 spargana were obtained from 20 frogs Hoplobatrachus rugulosus; syn: Rana rugulosa (Wiegmann, 1834) or R. tigrina (Steindachner, 1867). The plerocercoids were used for experimental infection of a dog. After 4 weeks of infection, an adult tapeworm was recovered from the intestine of the dog. Morphologically, the distinct features of Spirometra sp. (Myanmar origin) relative to S. erinaceieuropaei and S. decipiens include a uterine morphology comprising posterior uterine coils that larger than the terminal uterine ball and coiling of the uteri diagonally (swirling) rather than spirally. The cox1 sequences (1,566 bp) of the Myanmar-origin Spirometra species showed 97.9% similarity to a reference sequence of S. decipiens (GenBank no. KJ599679) and 90.5% similarity to a reference sequence of S. erinaceieuropaei (GenBank no. KJ599680). Phylogenetic tree topologies were identical and presented high confidence level of values for the 3 major branches of the 3 Spirometra species in cox1 and nad1 genes. These results indicated that Myanmar-origin Spirometra species coincided with those of S. ranarum and may be considered as a valid species.

Quali-Quantitative Analysis of Flavonoids for Mulberry Leaf and Fruit of 'Suhyang'

  • Ju, Wan-Taek;Kwon, O-Chul;Lee, Min-Ki;Kim, Hyun-Bok;Sung, Gyoo-Byung;Kim, Yong-Soon
    • Korean Journal of Environmental Agriculture
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    • v.36 no.4
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    • pp.249-255
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    • 2017
  • BACKGROUND: Globally, mulberry (Morus sp.) is exploited for feeding leaf to silkworms in order to obtain silk fiber or for animal feedstock production. Also, mulberry fruit is known to a by-product that was produced from mulberry tree after harvesting leaves for silkworm rearing, as a yield and consumption of mulberry fruit was increased, it has been fixing to a newincome crop. Mulberry leaves and fruits are used for the health benefits of human beings. Mulberry contains various bioactive components, such as alkaloids and flavonoids. Mulberry flavonoids are an important part of the diet because of their effects on human nutrition. The flavonoids in mulberry leaf and fruit of 'Suhyang'(Morus alba L.) were determined. METHODS AND RESULTS: Flavonoids for mulberry leaf and fruit of 'Suhyang' were analysed using ultrahigh performance liquid chromatography coupled with diode array detection and quadrupole time-of-flight mass spectrometry (UPLC-DAD-QTOF/MS)technique. An UPLC-DAD-QTOF/MS system was used, and identification of mulberry leaves constituents was carried out on the basis of the complementary information obtained from LC spectra, MS ions, and MS/MS fragments. The mulberry leaf (16 flavonoids) and fruit (9 flavonoids) were isolated and analyzed from Suhyang using UPLC-DAD-QTOF/MS chromatogram. To the best of our knowledge, Quercetin 3-O-(6"-O-malonyl) glucoside and quercetin 3-O-rutinoside (rutin) was detected on the highest content in leaf and fruit, respectively and further research will be devoted to evaluate their biological activity. CONCLUSION: Obtaining information about the concentration of functional materials in mulberry leaves could contribute to the development and promotion of processed, functional products and offer possible industrial use of 'Suhyang', holding promises to enhance the overall profitability of sericulture.

Analytical Characteristics of Electron Spin Resonance for Identifying Irradiated Ramen Soup with Radiation Sources (방사선 조사된 라면수프의 조사선원에 따른 전자스핀공명 분석특성)

  • Ahn, Jae-Jun;Lee, Ju-Woon;Chung, Hyung-Wook;Kwon, Joong-Ho
    • Korean Journal of Food Science and Technology
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    • v.41 no.2
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    • pp.131-135
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    • 2009
  • The identification characteristics of irradiated commercial Ramen soup were investigated depending on radiation sources and doses by electron spin resonance (ESR) spectroscopy. Two commercial powder soups (RS-1, RS-2) were irradiated at 0 to 20 kGy under ambient conditions by both a Co-60 gamma irradiator and an electron beam (EB) accelerator, respectively. Crystalline sugar-induced multi-component signals with g-values of 2.010/2.011, 2.006, 2.002 and 1.999 were detected in the irradiated Ramen soup (RS-1, RS-2), whereas $Mn^{2+}$ signals were observed in non-irradiated samples, thereby distinguishing each other. Under the same analytical conditions, the intensity of ESR signals was higher in EB-irradiated samples than the gamma-irradiated ones. Determination coefficients ($R^2$) between irradiation doses and corresponding ESR responses were above 0.9665 in all the samples, and the magnetic field of specified g-value remained constant. The predominant ESR signals of $g_2$ (2.010-2.011) and $g_3$ (2.002) increased with corresponding doses of irradiation ($R^2$= 0.9750-0.9981).

Molecular Cloning and Expression of Heat-stable Enterotoxin Gene from Swine Enterotoxigenic Escherichia coli (돼지에서 유래한 병원성 대장균의 내열성 장독소 생산유전자의 Cloning 및 발현)

  • 김교창;도대흥
    • Journal of Food Hygiene and Safety
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    • v.6 no.3
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    • pp.147-155
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    • 1991
  • Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenlc E. coli was isolated from swine during 1989 year(from 5 to 10 month) in the Kyong-gi and Chung-Cheong provinces, and three strains(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5~9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase was at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing strain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plasmid. Each fragment of this plasmid digested with EcoRl was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRl digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.

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