• 제목/요약/키워드: multiplex-PCR

검색결과 493건 처리시간 0.029초

Distribution of EGFR Mutations Commonly Observed in Primary Lung Adenocarcinomas in Pakistan as Predictors for Targeted Therapy

  • Ahmed, Zeeshan Ansar;Moatter, Tariq;Siddiqui, Areeba;Pervez, Shahid
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권17호
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    • pp.7125-7128
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    • 2014
  • Background: Acquired genetic alterations and presence of sensitizing mutations in the tyrosine kinase domain of EGFR and other signaling molecules have been found in different subsets of primary lung adenocarcinoma. The commonest EGFR mutations are small in frame deletions of exon 19 and a point mutation (L858R) in exon 21, having a combined occurrence of around 90%. The objective of this study was to determine the frequency and types of EGFR mutations in primary lung adenocarcinomas in Pakistan. Materials and Methods: EGFR mutations in tumor samples were screened by multiplex real time PCR. Briefly, DNA from formalin fixed paraffin-embedded tissue was amplified with primers and probes specific to 43 different EGFR mutations in a Cobas z 480 instrument. The assay detects mutations in four exons (18-21) of the EGFR gene. Results: Out of 94 patients, 65 were males and 29 females with a M:F ratio of 2.2: 1. The median age was 62 years (range, 28 - 85 years). In our biopsy samples 70 (74%) cases were of primary lung adenocarcinoma, whereas 24 (26%) were confirmed metastatic adenocarcinoma of primary lung origin. EGFR mutation was positive in 29% of the patients. The highest frequency of L858R was observed in 48% of these, followed by deletion in exon 19 (44%). In addition, other rare mutations such as compound G718X:S768I and insertions in exon 20 insertion were detected in approximately 4% of the patients. Conclusions: This study showed that Del 19 and L858R are the most frequent mutations in Pakistani lung adenocarcinoma patients and around 29% of the patients were found eligible for erlotinib therapy.

Comparative study of Clostridium perfringens, Salmonella spp. and E. coli focused on characteristics of E. coli O157 isolated from pigs of HACCP- and non-HACCP-accredited swine farms in Korea

  • Keum, Hyun Ok;Kim, Hye Kwon;Rho, Se Mi;Moon, Hyoung Joon;Park, Seong Jun;Park, Bong Kyun
    • 대한수의학회지
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    • 제50권2호
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    • pp.113-116
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    • 2010
  • To determine the prevalence of Escherichia (E.) coli O157 : H7 from pigs after the Hazard Analysis and Critical Control Point (HACCP) system has been applied to Korean swine farm since 2006, 291 fecal samples were tested between May and December in 2008. Four E. coli O157:non-H7 (1.4%) were isolated from 4 different non-HACCP-accredited farms and they didn't have virulent genes which can cause illness for human. Also, Clostridium (C.) perfringens, Salmonella spp. and E. coli enterotoxins were tested using multiplex PCR. The positive rate for these pathogens of non-HACCP-accredited farms was higher than that of HACCP-accredited farms, and especially in case of C. perfringens, E. coli enterotoxins LT and STa, it was statistically significant (p < 0.05). Thus, the early implementation of the HACCP program is expected to greatly contribute to the safety of livestock products as well as food hygiene.

주거환경 공기 중 호흡기 바이러스의 검출 (Detection of Airborne Respiratory Viruses in Residential Environments)

  • 박근태;문경환;김형태;박찬정;정호철;임영희
    • 한국환경보건학회지
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    • 제37권4호
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    • pp.306-314
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    • 2011
  • Objectives: Respiratory virus infections are the most common disease among all ages in all parts of the world and occur through airborne transmission. The purpose of this study was to detect and quantitate human respiratory viruses in residential environments. Methods: Air samples were collected from the residential space of apartments in the Seoul/Gyeonggi-do area. The samples were collected from indoor and outdoor air. Among respiratory viruses, influenza A virus, influenza B virus, parainfluenza virus, metapneumovirus, respiratory syncytial virus, and adenovirus were investigated by multiplex polymerase chain reaction. Among the virus-positive samples, we performed adenovirus quantification by real-time polymerase chain reaction. Results: Virus detection rates were 44.0%, 3.8%, 3.4%, and 17.3% in spring, summer, autumn, and winter, respectively. The virus detection rate was higher in winter and spring than in summer and autumn. Adenovirus was most commonly detected, followed by influenza A virus and parainfluenza virus. Virus distribution was not significantly different between indoor and outdoor environments. Conclusions: Although virus concentrations were not high in residential environments, residents in houses with detected viruses may have an increased risk of exposure to airborne respiratory viruses, especially in winter and spring.

Glutathione S-Transferase T1 and M1 Polymorphisms and Risk of Uterine Cervical Lesions in Women from Central Serbia

  • Stosic, Ivana;Grujicic, Darko;Arsenijevic, Slobodan;Brkic, Marija;Milosevic-Djordjevic, Olivera
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권7호
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    • pp.3201-3205
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    • 2014
  • The aim of this study was to investigate the frequencies of GSTT1 and GSTM1 deletion polymorphisms in newly-diagnosed patients with uterine cervical lesions from central Serbia. Polymorphisms of GST genes were genotyped in 97 patients with cervical lesions and 50 healthy women using a multiplex polymerase chain reaction (PCR). The GSTM1 null genotype was significantly more prominent among the patients than in controls (74.2% vs 56.0%), the risk associated with lesions being almost 2.3-fold increased (OR=2.26, 95%CI=1.10-4.65, p=0.03) and 3.17-fold higher in patients above >45 years old (95%CI=1.02-9.79, p=0.04). The analysis of the two genotypes demonstrated that GSTM1 null genotype significantly increased risk only for low grade squamous intraepithelial lesion-LSIL (OR=2.81, 95%CI=1.03-7.68, p=0.04). GSTT1 null genotype or different genotype combinations were not found to be risk factors, irrespective to lesion stages, age or smoking. We found that the risk of cervical lesions might be significantly related to the GSTM1 null genotype, especially in women aged above 45 years. Furthermore, the GSTM1 polymorphism might have greater role in development of early stage lesions.

Increase in the Rate of HPV Positive Oropharyngeal Cancers During 1996-2011 in a Case Study in Turkey

  • Tural, Deniz;Elicin, Olgun;Batur, Sebnem;Arslan, Deniz;Oz, Buge;Serdengecti, Suheyla;Uzel, Omer
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권10호
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    • pp.6065-6068
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    • 2013
  • Background: Primary aim of this study is to assess whether or not there is an increase at rate of HPV positive oropharyngeal cancers during 1996-2011 in Turkey, for comparison with prior reports from Western countries. Materials and Methods: A total of 138 newly diagnosed patients with oropharyngeal cancer were identified, 39 of which had no primary tumor specimen available and 18 patients with invalid HPV status, therefore HPV status for remaining 81 patients was evaluated. The presence and type of HPV DNA were determined with formalin-fixed paraffin embedded specimens, using an HPV DNA-based multiplex PCR assay. Associations between HPV status and clinicopathological characteristics were evaluated using a two-sample t-test for the continuous variables and the categorical variables were compared by chi-square test. Overall survival (OS) periods were calculated with Kaplan-Meier method. Results: The proportion of HPV-positive cancer has continued to increase during 2004-2011 as compared with 1996-2003. Notably, 33% (6/18) of the cases were HPV-positive in 1996-1999, 43% (9/21) in 2000-2003, 55% (11/20) in 2004-2007 and 70% (16/23) in 2008-2011. Thus, when we compared the results obtained during the 2004-2011with results of 1996-2003 period, we found that increase at HPV-positivity ratio was statistically significant (38% vs 64% p=0.012). Conclusions: This study demonstrated that HPV positive oropharyngeal cancers are increasing in Turkish patients as in the Western world.

Candida non albicans with a High Amphotericin B Resistance Pattern Causing Candidemia among Cancer Patients

  • Kalantar, Enayatollah;Assadi, Mojan;Pormazaheri, Helen;Hatami, Shiva;Barari, Maryam Agha;Asgari, Esfandiar;Mahmoudi, Elaheh;Kabir, Kourosh;Marashi, Seyed Mahmoud Amin
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권24호
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    • pp.10933-10935
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    • 2015
  • Background: Many scientists have reported Candida species to be of great concern because of the high frequency that they colonize and infect human hosts, particularly cancer patients. Moreover, in the last decades Candida species have developed resistance to many antifungal agents. Based on this, we aimed to identify and determine the prevalence of Candida spp from blood culture bottles among cancer patients and their antifungal resistance pattern. Materials and Methods: From the blood culture bottles isolation and identification of the Candida spp were performed by conventional microbiological techniques. The in vitro antibiotic resistance pattern of the isolates was determined by CLSI guidelines. Genomic DNA was isolated and amplified. Each gene was separated by agar gel electrophoresis. Results: Identification of Candida spp was based on the presence of yeast cells in direct examination, culture and DNA extraction. Of the 68 blood samples collected during the study period (April 2013 to October 2013), five (7.35%) were positive for the presence of Candida spp, 2 (40%) of which were identified as Candida albicans and 3 (60%) were Candida non-albicans. Conclusions: High resistance to amphotricin B was observed among all the Candida non-albicans isolates. Regular investigations into antifungal resistance will help us to get an updated knowledge about their antibiotic resistance pattern which may help the physician in selecting the antibiotics for empirical therapy.

Laboratory Investigation of Trends in Bacterial Pneumonia in Cheonan, Korea, from January 2008 to September 2017

  • Yook, Young-Sam;Jeon, Jae-Sik;Park, Ji On;Kim, Jae Kyung
    • Journal of Microbiology and Biotechnology
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    • 제28권10호
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    • pp.1730-1735
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    • 2018
  • Bacterial pneumonia is one of the most common causes of mortality in Korea. In 2016, the mortality rate from pneumonia was 16,476 deaths per 100,000, which was an 11% increase from the previous year. The aim of our study was to determine the distribution of the bacterial pathogens causing respiratory symptoms in different age groups over a 10-year period. Between January 2008 and September 2017, 1,861 specimens from 1,664 patients admitted to Dankook University Hospital with respiratory symptoms were examined. We used multiplex polymerase chain reaction (PCR) to detect six bacterial pneumonia pathogens: Bordetella pertussis, Chlamydophila pneumoniae, Haemophilus influenzae, Legionella pneumophila, Mycoplasma pneumoniae, and Streptococcus pneumoniae. We detected bacterial pneumonia pathogens in 1,281 (68.83%) specimens. Of the 1,709 pathogens detected, S. pneumoniae was the most common (48.57%; n = 830) followed by H. influenzae (40.08%; n = 685). Most infections were found among children younger than 10 years (92.69%; n = 1,584). Although S. pneumoniae was the most common pathogen detected in all age groups, M. pneumoniae infection increased in prevalence with age (p < 0.05). The rate of co-infection was also high among these patients (31.1%; n = 399), which peaked in 2015 (54.55%; n = 42/77). The prevalence of bacterial pneumonia in Cheonan, along with the proportion of co-infections among patients increased over the 10-year study period. The findings will aid the development of treatment and prevention guidelines.

Forensic Characterization of Four New Bovine Tri-nucleotide Microsatellite Markers in Korean Cattle (Hanwoo)

  • Sim, Yong Teak;Na, Jong Gil;Lee, Chul-Sang
    • Journal of Animal Science and Technology
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    • 제55권2호
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    • pp.87-93
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    • 2013
  • We identified four new bovine tri-nucleotide microsatellite loci and analyzed their sequence structures and genetic parameters in 105 randomly selected Korean cattle (Hanwoo). Allele numbers of the loci B17S0808, B15S6253, B8S7996, and B17S4998 were 10, 11, 12, and 29, respectively. These alleles contained a simple or compound repeat sequences with some variations. Allele distributions of all these loci were in Hardy-Weinberg equilibrium (P > 0.05). Observed heterozygosity and expected heterozygosity ranged from 0.54 (B15S6253) to 0.92 (B17S4998) and from 0.599 (B15S6253) to 0.968 (B17S4998), respectively, and two measures of heterozygosity at each locus were highly correlated. Polymorphism information content (PIC) for these 4 loci ranged from 0.551 (B15S6253) to 0.932 (B17S4998), which means that all these loci are highly informative (PIC > 0.5). Other genetic parameters, power of discrimination (PD) and probability of exclusion (PE) ranged from 0.783 (B15S6253) to 0.984 (B17S4998) and from 0.210 (B15S6253) to 0.782 (B17S4998), respectively. Their combined PD and PE values were 0.9999968 and 0.98005176, respectively. Capillary electrophoresis revealed that average peak height ratio for a stutter was 13.89% at B17S0808, 26.67% at B15S6253, 9.09% at B8S7996, and 43.75% at B17S4998. Although the degree of genetic variability of the locus B15S6253 was relatively low among these four microsatellite markers, their favorable parameters and low peak height ratios for stutters indicate that these four new tri-nucleotide microsatellite loci could be useful multiplex PCR markers for the forensic and population genetic studies in cattle including Korean native breed.

Coexistence of Malaria and Thalassemia in Malaria Endemic Areas of Thailand

  • Kuesap, Jiraporn;Chaijaroenkul, W.;Rungsihirunrat, K.;Pongjantharasatien, K.;Na-Bangchang, Kesara
    • Parasites, Hosts and Diseases
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    • 제53권3호
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    • pp.265-270
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    • 2015
  • Hemoglobinopathy and malaria are commonly found worldwide particularly in malaria endemic areas. Thalassemia, the alteration of globin chain synthesis, has been reported to confer resistance against malaria. The prevalence of thalassemia was investigated in 101 malaria patients with Plasmodium falciparum and Plasmodium vivax along the Thai-Myanmar border to examine protective effect of thalassemia against severe malaria. Hemoglobin typing was performed using low pressure liquid chromatography (LPLC) and ${\alpha}$-thalassemia was confirmed by multiplex PCR. Five types of thalassemia were observed in malaria patients. The 2 major types of thalassemia were Hb E (18.8%) and ${\alpha}$-thalassemia-2 (11.9%). There was no association between thalassemia hemoglobinopathy and malaria parasitemia, an indicator of malaria disease severity. Thalassemia had no significant association with P. vivax infection, but the parasitemia in patients with coexistence of P. vivax and thalassemia was about 2-3 times lower than those with coexistence of P. falciparum and thalassemia and malaria without thalassemia. Furthermore, the parasitemia of P. vivax in patients with coexistence of Hb E showed lower value than coexistence with other types of thalassemia and malaria without coexistence. Parasitemia, hemoglobin, and hematocrit values in patients with coexistence of thalassemia other than Hb E were significantly lower than those without coexistence of thalassemia. Furthermore, parasitemia with coexistence of Hb E were 2 times lower than those with coexistence of thalassemia other than Hb E. In conclusion, the results may, at least in part, support the protective effect of thalassemia on the development of hyperparasitemia and severe anemia in malaria patients.

사상체질별 Short Tandem Repeat 대립유전자 빈도 (Short Tandem Repeat Allele Frequencies in Sasang Constitution)

  • 박화용;유현주;구임회;김종열
    • 사상체질의학회지
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    • 제21권1호
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    • pp.227-236
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    • 2009
  • 1. Objectives As a basic trial for identification of Sasang constitutional gene marker, we genotyped and analysed statistical relationships of STR(short tandem repeat) alleles and its distribution in each constitution. 2. Methods After obtaining basic constitutional data with questionnaire (QSCC II), decision of constitution was made by 3 different constitution specialists' diagnosis, and only the samples of specialists' agreement of each constitution by discussion were taken into this research. Using multiplex PCR kit, total 146 constitutional samples were amplified in 16 autosomal STR marker, genotyped, and analysed statistically. Among 16 markers, 15 were analysed in this study excluding the amelogenin marker is used for in gender identification. 3. Results and Conclusions It is difficult to determine the relationship between constitution and STR marker as the sample size is small, however, Penta D and vWA were shown to be related statistically with constitution. It has been know that STRs has no genetic informations, however there are some recent research results showing STRs as a regulatory element, relationship between microsatellite instability and repeat number and size, and post-transcriptional sigualing. STRs which is not known about its function currently, are proposed to have function and/or regulatory activities anyhow with Sasang constitution. It is believed that the results of this study can halp determine and deatify the markers related to Sasang Constitutional Medicien.

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