• Title/Summary/Keyword: TaqI

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Development of TaqMan Quantitative PCR Assays for Duplex Detection of Dirofilaria immitis COI and Dog GAPDH from Infected Dog Blood (심장사상충에 감염된 개 혈액에서 Dirofilaria immitis의 COI와 개의 GAPDH를 이중 검출하기 위한 정량적 TaqMan PCR 분석법의 개발)

  • Oh, In Young;Kim, Kyung Tae;Gwon, Sun-Yeong;Sung, Ho Joong
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.1
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    • pp.64-70
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    • 2019
  • Dirofilaria immitis (D. immitis) is a filarial nematode that causes cardiopulmonary dirofilariasis in dogs. In the late stages of infection, infected dogs show one or more symptoms and advanced heart disorder with perivascular inflammation. To detect D. immitis specifically and efficiently in the early stages of infection, a duplex TaqMan qPCR assay was developed based on previous studies using primers and probes specialized to detect D. immitis cytochrome c oxidase subunit I (COI) and dog glyceraldehyde-3-phosphate dehydrogenase (GAPDH). As positive controls, plasmid DNAs were constructed from D. immitis COI or dog GAPDH and a TA-cloning vector. Simplex and duplex TaqMan qPCR assays were performed using the specific primers, probes, and genomic or plasmid DNA. The duplex reaction developed could detect D. immitis COI and dog GAPDH in the same sample simultaneously after optimization of the primer concentrations. The limit of detection was 25 copies for the simplex and duplex assays, and both showed good linearity, high sensitivity, and excellent PCR efficiency. The duplex assays for pathogen detection reduce the costs, labor, and time compared to simplex reactions. Therefore, the duplex TaqMan qPCR assay developed herein will allow efficient D. immitis detection and quantification from a large number of samples simultaneously.

A Study on Sasanian Costume Depicted on Rock Reliefs at Taq-I Bustan (타퀴이 부스탄 부조에 묘사된 사산조 복식연구)

  • Youngsoo Chang
    • Journal of Fashion Business
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    • v.27 no.2
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    • pp.1-25
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    • 2023
  • Taq-i Bustan rock reliefs are relatively well preserved among Sassanid relics. They are considered concrete and empirically important materials for studying Sassanian culture. In these rock reliefs, inauguration of various kings and hunting scenes are depicted. Thus, kings' attire and life of the court could be discerned according to eras. The purpose of this study was to investigate Sassanian clothing styles of various social positions by analyzing their clothing depicted in rock reliefs and to improve comprehension of the culture of the Sassanian Dynasty. Methodically, literature studies and artifacts were analyzed in parallel. Data of these two insources were consolidated for comparison. Results of this study are as follows. Since the Sassanian Dynasty succeeded the Parthian Dynasty, the Parthian style of clothing pieces such as tunic and trousers appeared in their attire. Basic types of tunics and trousers did not vary according to social status or age. However, details showed differences in those aspects. In the early Sassanian Dynasty, the dressing style of Rome was adopted. Styles of tunic and trousers featured draperies with many wrinkles and shoes were worn instead of boots. Trousers were tied with laces at ankles and laces were decorated with round clasps, imitating the style used by Kushan kings to the east of Sasanians. External cultural elements were also present in costumes of the Sassanian Dynasty, which played a bridging role for cultural exchanges between Eastern and Western civilizations at the gateway of the Silk Road.

Association of Vitamin D Receptor Gene Polymorphisms with Prostate Cancer Risk in the Pakistani Population

  • Yousaf, Nageen;Afzal, Sibtain;Hayat, Tehreem;Shah, Jasmin;Ahmad, Nafees;Abbasi, Rashda;Ramzan, Khushnooda;Jan, Rasul;Khan, Imran;Ahmed, Jawad;Siraj, Sami
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.10009-10013
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    • 2014
  • Background: Vitamin D receptor (VDR) gene has been a subject of extensive pharmacogenetic research recently. Association studies between different types of cancers including prostate cancer (PCa) and VDR gene polymorphism have also been conducted. The objective of this study was to find possible associations between PCa and VDR gene polymorphisms in the Pakistani population. Materials and Methods: A total of 162 subjects, including prostate cancer patients and controls, were genotyped for Apa I, Taq I and Fok I polymorphisms in the VDR gene using allele specific PCR, PCR-RFLP and direct DNA sequencing. Allelic frequencies were tested for Hardy-Weinberg equilibrium and associations between the genetic markers and PCa were calculated using logistic regression. Results: Apa I CC genotype was found to have strongest association with PCa risk, and "A" genotype was found to have protective effect. Fok I and Taq I did not have appreciable levels of association with PCa, although Taq I "TC" heterozygotes seemed to have some protective effect. Similarly the "C" allele of Fok I also seemed to have protective effect. Conclusions: To our knowledge, this is the first report showing association between VDR gene polymorphisms and PCa in Pakistan. Our findings may be somewhat skewed because of small sample size and tendency of consanguineous marriages in Pakistani society; nevertheless, it shows the trend of association and protective effects of certain VDR gene polymorphisms against PCa.

A Simple Method for Elimination of False Positive Results in RT-PCR

  • Martel, Fatima;Grundemann, Dirk;Schomig, Edgar
    • BMB Reports
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    • v.35 no.2
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    • pp.248-250
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    • 2002
  • Discrimination between the amplification of mRNA and contaminating genomic DNA is a common problem when performing a reverse transcriptase-polymerase chain reaction (RT-PCR). Even after treatment of the samples with DNAse, it is possible that negative controls (samples in which no reverse transcriptase was added) will give positive results. This indicates that there was amplification of DNA, which was not generated during the reverse transcriptase step. The possibility exists that Taq DNA polymerase acts as a reverse transcriptase, generating cDNA from RNA during the PCR step. In order to test this hypothesis, we incubated samples with a DNAse-free RNAse after the cDNA synthesis. Comparison of the results that were obtained from these samples (incubated with or without DNAse-free RNAse) confirms that the reverse transcriptase activity of Taq DNA polymerase I is a possible source of false positive results when performing RT-PCR from intronless genes. Moreover, we describe here a simple and rapid method to overcome the false positive results that originate by this activity of Taq polymerase.

A FAMILY-BASED AND CASE-CONTROL ASSOCIATION STUDY OF THE DOPAMINE BETA HYDROXYLASE GENE POLYMORPHISM IN ATTENTION DEFICIT HYPERACTIVITY DISORDER (한국인 주의력결핍 과잉행동장애와 Dopamine Beta Hydroxylase 유전자의 관련성 : 가족기반 연구 및 환자-대조군 연구)

  • Park Tae Won;Kim Boong Nyun;Im Myung-Ho;Yoo Hee Jeone;Kang Daehee;Cho Soo Churl
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.16 no.1
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    • pp.54-62
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    • 2005
  • Objective : Attention deficit hyperactivity disorder (ADHD) is the most common childhood psychiatric disorder, affecting $3-5\%$ of school age children. Although the biological basis of ADHD is unknown, family studies provide strong evidence that ADHD has a genetic basis. Recent genetic studies have suggested associations between ADHD and Taq I polymorphism of dopamine beta hydroxylase gene(DBH) . The aim of this study is to test the association between ADHD and Taq I polymorphism of DBH in Korean population. Method : We processed DNA extraction and genotyping for 106 korean children with ADHD and their parents. Genotyping was additionally performed for 212 age and gender matched normal controls. Case-control association study was applied. And we tested the association using the transmission disequilibrium test (TDT) and haplotype-based haplotype relative risk test (HHRR). Results : There were no statistical differences of genotype distributions between cases and controls. However, we did observe preferential transmission of allele Al of DBH Taq T polymorphism in ADHD. Conclusion : On the whole, our results lend credence to the notion that the relationship between ADHD and DBH is complex. The number of cases and informative transmissions were small, therefore it would be premature to make any conclusions from our study concerning the role of DBH in ADHD. Further work is needed to support these findings.

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DNA Analysis of Ginseng Using PCR-aided RFLP Technology (PCR-aided RFLP기술을 이용한 인삼의 DNA분석)

  • Yang, Deok-Chun;Kim, Moo-Sung
    • Journal of Ginseng Research
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    • v.27 no.3
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    • pp.146-150
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    • 2003
  • This study was carried out to obtain basic information on breeding using PCR-aided RFLP technology which can identify the variation inter- and intra-species of ginseng in the level of DNA. It was intended to investigate banding pattern on psbA and rbeL genes of chloroplast DNA in ginseng after treating with restriction enzymes. To isolate psbA and rbcL genes of chloroplast, both psbA-N, psbA-C primer and rbcL-N, PX-1 primer were used. As a result, 1,008 bp band of psbA gene and 1,336 bp band of rbcL gene were appeared, which was optimal and expected molecular weight. In addition, primers to isolate atpB, rpoB, trnL, and trnF genes were used, resulting in the expected 1366, 900, 1500 and 1008 bp bands. Genes of psbA and rbcL isolated by PCR were cut by restriction enzymes, Sau3A, TaqI, AluI, HaeIII, and RFLP pattern was investigated. KG line and other species of ginseng were cut by TaqI treatment, and bands were located in 800 bp. The treatment treated by AluI also showed the same 800 bp band in KG line and other species. In HaeIII treatment, 500 bp of faint bands were shown in case of KG line, whereas any bands were not observed in other species. All chloroplast genes formed bands by PCR amplification. However, it was not evident to distinguish intra-or inter-species of ginseng after restriction enzyme treatment. Therefore, more restriction enzyme treatment or sequence comparison method should be considered for further experiment.

Development of Real-time PCR Assays for Detection of Dirofilaria immitis from Infected Dog Blood (심장사상충에 감염된 개의 혈액에서 심장사상충 유전자를 검출할 수 있는 실시간 중합효소연쇄반응 기법 개발)

  • Oh, In Young;Kim, Kyung Tae;Jun, Jin Hyun;Shin, Jae-Ho;Sung, Ho Joong
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.88-93
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    • 2016
  • Dirofilaria immitis is a filarial nematode parasite that causes cardiopulmonary dirofilariasis in dogs. The purpose of this study was the development of real-time PCR assays for efficient detection of D. immitis. The D. immitis-specific primers confirmed in our previous study and a newly designed TaqMan probe were used for quantitative diagnostics. First, SYBR Green real-time PCR was performed using the specific primers and serially diluted genomic DNA or plasmid DNA, and melting curve analyses were performed after amplification. The melting curve showed one specific peak in each of the genomic and plasmid DNA reactions, suggesting that the primers specifically amplify the D. immitis cytochrome c oxidase subunit I gene. Comparison of SYBR Green and TaqMan real-time PCR using serially diluted plasmid DNA showed higher efficiency and specificity with TaqMan real-time PCR. The real-time PCR assays developed in this study will provide improved diagnostic methods to overcome the limitations of conventional diagnostic tools and facilitate more rapid and accurate diagnoses.

Association Study between Tic Disorder and Dopamine D2 Receptor Gene Polymorphism in Korean Population (틱장애와 도파민 D2 수용체 유전자와의 연합 연구)

  • Lee, Soyoung Irene;Cho, In Hee;Kim, Seon Mee;Lee, Min-Soo;Jung, Han-Yong
    • Korean Journal of Biological Psychiatry
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    • v.13 no.4
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    • pp.299-304
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    • 2006
  • Objectives : The purpose of the present study was to investigate whether the TaqI A polymorphism of dopamine receptor D2 gene(DRD2) is associated with Tourette syndrome(TS) and chronic motor tic disorder(CMT) in Korean population. Methods : DRD2 TaqI A RFLP genotyping was carried out with DNA extracted from blood samples of 75 patients with tic disorders(47 with TS and 28 with CMT) and 90 healthy subjects. Genotype and allelic frequencies for the DRD2 gene polymorphisms of the tic disorder group as a whole were compared to those of the control group. Separating the TS group, thereafter, the frequency of genotypes and alleles were compared to those of the controls. Results : The results demonstrated that genotype and allele distributions for the DRD2 gene polymorphism in the tic disorder as a whole, TS, and control groups were not significantly different. Conclusion : No association was found for DRD2 gene, TS and CMT. The data suggest that DRD2 gene may not be a useful marker for the prediction of the susceptibility of tic disorder.

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NEONATAL BEHAVIORAL CHARACTERISTICS AND DOPAMINE TRANSPORTER GENE AND DOPAMINE D2, D3, D4 RECEPTOR GENE POLYMORPHISMS (신생아 행동 특성과 Dopamine Transporter 유전자 및 Dopamine D2, D3, D4 수용체 유전자의 다형성)

  • Park, Young-Nam;Kim, Dae-Kwang;Kim, Sung-Wook
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.12 no.2
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    • pp.179-191
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    • 2001
  • Objectives:This study evaluated the association between behavioral characteristics and polymorphisms in DAT1, DRD2, DRD3, and DRD4 genes. Methods:The subjects were 114 neonates, who were born by normal spontaneous vaginal delivery and had no physical problems. The behavioral characteristics were evaluated using Neonatal Behavioral Assessment Scale(NBAS) at $17.8{\pm}7.0$ hours after their birth to minimize environmental influences, and cord blood was used to analyze the gene polymorphisms. Results:In comparison to DAT1 gene 10/10 genotype group(N=93), other genotype group(N=19) showed significantly high NBAS scores on social-interaction, state organization, and state regulation. DRD2 gene Ser311/Cys311, TaqI A, and TaqI B polymorphisms showed no significant differences on NBAS scores when they were grouped by genotypes. DRD3 gene polymorphism and DRD4 gene promotor polymorphism showed no significant difference on NBAS scores when they were grouped by genotypes. In comparison to the short repeats(N=102), long repeats(N=10) in DRD4 gene showed significantly high habituation score of NBAS. Conclusion:These results suggest that the genetic effects of the neonatal behavioral characteristics may be mediated via DAT1 and DRD4 genes.

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D2 Dopamine Receptor (DRD2) Gene Polymorphism and Combat-Related Posttraumatic Stress Disorder in Vietnam Veterans (월남전 참전 재향군인에서 도파민 D2 수용체 유전자 다형성과 외상후 스트레스 장애)

  • Lee, Soo-Young;Chung, Hae-Gyung;Kim, Tae-Yong;Choi, Jin-Hee;Chung, Moon-Yong;So, Hyoung-Seok;Shin, Han-Sang;Lee, Shi-Eun
    • Anxiety and mood
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    • v.4 no.2
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    • pp.142-147
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    • 2008
  • Objective : Evidence from recent studies supports the role of genetic factors in the development of Posttraumatic Stress Disorder (PTSD). The primary aim of this study is to investigate the association between the dopamine D2 receptor (DRD2) TaqI A polymorphism and PTSD. The second aim is to examine the association between the DRD2 TaqI A polymorphism and clinical symptoms in patients with PTSD. Methods : We recruited 189 Vietnam veterans for participation in this study, among whom 99 were PTSD patients and 90 were control subjects. The presence of the DRD2 TaqI A polymorphism was determined by polymerase chain reaction (PCR). Several standardized research scales were used in the clinical assessment of PTSD, including the Combat Exposure Scale (CES), Clinician Administered PTSD Scale (CAPS), Beck Depression Inventory (BDI), and Clinical Global Impression (CGI). Results : There was no significant difference in the distribution of the DRD2 genotype, frequency and prevalence of the A1 allele, or the frequency of heterozygotes between the patients with PTSD and the controls. In the PTSD group, the patients with the A1 allele (A1A1, A1A2) scored higher on the CAPS-total (p=0.044), CAPS-avoidance symptoms (p=0.016) and BDI (p=0.024) than those without the A1 allele (A2A2). Conclusion : We could not find an association between the dopamine D2 receptor (DRD2) TaqI A polymorphism and PTSD. However, the A1 allele of DRD2 seemsto influence avoidance symptoms in patients with PTSD.

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