• Title/Summary/Keyword: DNA screening

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Evaluation of Several Screening Approaches for Detection of Cervical Lesions in Rural Shandong, China

  • Zong, Li-Ju;Zhang, You-Zhong;Yang, Xing-sheng;Jiang, Jie;Cui, Bao-Xia;Qiao, Yun-Bo;Li, Li;Jiang, Kan;Zhang, Wen-Jing;Kong, Bei-Hua;Shen, Keng
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.1907-1912
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    • 2015
  • Purpose: The study was designed to: (1) investigate the prevalence of high-risk human papillomavirus (HR-HPV) infection and cervical neoplasia; and (2) evaluate clinical performance of visual inspection with acetic acid/ Lugol's iodine (VIA /VILI), Pap smear, high-risk human papillomavirus (HR-HPV) DNA test for detecting cervical intraepithelial neoplasia grade 2 or worse (CIN2+) and (3) explore appropriate screening approach in rural areas of Shandong Province. Materials and Methods: A total of 3,763 eligible women from Yiyuan County in Yimeng mountainous areas of rural Shandong, China, were enrolled and underwent Pap smear, HR-HPV DNA testing by Hybrid Capture 2 (HC2), and VIA /VILI tests. Women positive in any test were referred to colposcopy and biopsy as indicated. Results: The prevalence of HR-HPV infection among all enrolled women was 11.1% and that in healthy women was 9.9%. In total 33 cases of CIN1, 16 cases of CIN2, 6 cases of CIN3 but none of cervical cancer were detected and the crude prevalence of CIN2+ was 0.58%. For detecting CIN2+, the sensitivity of HR-HPV DNA testing, VIA/VILI, Pap smear was 90.9%, 77.3%, 81.8%, respectively. Pap smear had the best specificity of 98.2%, followed by HR-HPV DNA testing with specificity of 89.4%, VIA/VILI had the lowest specificity of 81.2%. Colposcopy referral rate of HR-HPV DNA testing, VIA/VILI, Pap smear was 11.1%, 18.5%, 2.3%, respectively. Conclusions: Our results suggest that HR-HPV DNA testing alone might be appropriate for primary cervical cancer screening in rural low-resource areas of Shandong Province, China.

Experience of Combined Liquid Based Cervical Cytology and High-Risk HPV mRNA for Cervical Cancer Screening in Thammasat University Hospital

  • Muangto, Teerapat;Chanthasenanont, Athita;Lertvutivivat, Supapen;Nanthakomon, Tongta;Pongrojpaw, Densak;Bhamarapravatana, Kornkarn;Suwannarurk, Komsun
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.9
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    • pp.4409-4413
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    • 2016
  • Background: Cervical cancer is the second most common of malignancy found in Thai women. Human papillomavirus (HPV) infection is a major cause. The objective of the present study was to evaluate the prevalence of HPV infection and association with abnormal cervical cytology in Thai women. Materials and Methods: This study was conducted at the Gynecologic Clinic, Thammasat University, Pathum Thani, Thailand. A total of 2,144 cases who underwent annual cervical cancer screening by co-testing (liquid based cytology and HPV testing, DNA versus mRNA) during the priod from July 2013 to June 2016 were recruited in this study. Results: Prevalence of positive high risk (HR) HPV DNA and mRNA test were 19.7 and 8.4%, respectively with a statistically significant difference. Majority of cases of abnormal cytology in this study were atypical squamous cells of undetermined significance (ASC-US). In patients with ASC-US, positive HR HPV DNA was greater than in the mRNA group (10.1 and 4.5%, p<0.001). Nonetheless, there was no significant difference in participants with cervical intraepithelial neoplasia (CIN). HPV mRNA test had slightly lower sensitivity but higher negative predictive value (NPV) than the DNA test to detect abnormal cytology during cervical cancer screening (p<0.001). Both HPV test (DNA and mRNA) had equally efficacy to detect high grade precancerous lesion or higher (CIN 2+). Conclusions: Prevalence of HR HPV DNA and mRNA were 19.7 and 8.4 percent, respectively. NPV of HPV mRNA was higher than DNA test. Both tests had equal efficacy to detect CIN 2+ with sensitivity and specificity of 63% vs 55.7% and 83% vs 92%, respectively.

Evaluation of Methods to Analyze SNP-based Association Studies in a DNA-Pooling Experiment with Preferential Amplification

  • Ahn, Chul;Lee, Kyu-Sang
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.395-398
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    • 2005
  • Genetic association case-control studies using DNA pools are efficient ways of detecting association between a marker allele and disease status. DNA pooling is an efficient screening method for locating susceptibility genes associated with the disease. However, DNA pooling is efficient only when allele frequency estimation is done precisely and accurately. Through the evaluation of empirical type I errors and empirical powers by simulation, we will evaluate the methods that correct for preferential amplification of nucleotides when estimating the allele frequency of single-nucleotide polymorphisms.

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벼로부터 chloroplast small heat shock protein cDNA의 cloning 및 characterization

  • 이병현;원성혜;이효신;김기용;김미혜;정동민;조진기
    • Proceedings of the Korean Society of Grassland Science Conference
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    • 1999.06a
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    • pp.71.2-72
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    • 1999
  • 고등식물에 있어서 엽록체에 존재하는 저 분자량 heat shock protein (smHSP)은 식물의 내열성 획득에 있어서 필수유전자임이 mutant를 이용한 유전학적인 연구에 의해 보고된 바 있다. 고온내성이 강한 작물인 벼로부터 엽록체 smHSP cDNA를 분리하고자 벼의 잎에서 분리한 mRNA로 작성한 cDNA library로부터 screening하였다. 선발된 smHSP cDNA는 1,026 bp의 염기로 구성되어 있었으며, 239개의 아미노산으로 구성되는 예상분자량 26.6 kDa의 단백질을 code하고 있었다. 또한 다른 식물로부터(중략)

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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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    • v.35 no.4
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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.

Molecular cloning and characterization of metallothionein cDNA gene in channel catfish (챠넬메기의 metallothionein cDNA 유전자의 cloning 및 그 특성에 관한 연구)

  • Lee, In-Jung;Song, Young-Hwan
    • Journal of fish pathology
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    • v.5 no.2
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    • pp.143-152
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    • 1992
  • Metallothionein is an essential and common protein to regulate the intracellular concentration of heavy metals, which exist in most organisms from bacteria to vertebrates. Although the detailed function of metallothianein has not been fully identified until yet, it may be involoved in the cellular protection against the heavy metal toxicity and in the global regulation of several other genes and the expression of metalloproteins. We have cloned the full cDNA clone of metallothionein gene in Channel Catfish by Reverse Transcriptase-Polymerase Chain Reaction(RT-PCR) starting from poly(A)-containing mRNAs. All PCR fragments have been subcloned into EcoRV site of pBluescript SK+ and dT-tailed at Smal site of pUC19, then PCR products are recovered by the double digestion of recombinant plasmids wiht EcoRI and HindIII, which are adjacent to EcoRV site in multicloning sites or by rapid PCR screening. The nucleotide sequence analysis of pMT150(one of the PCR clones) showed high homology with several other piscine metallothionein cDNA genes.

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Cloning and Expression of Human Clotting Factor 9 cDNA un Escherichia coli (인체 혈액응고 9인자 cDNA cloning 및 Escherichia coli 에서의 발현)

  • Young Won Lee;Hyang Suk Hur;Myoung Hee Kim
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.231-240
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    • 1996
  • Human blood clotting (coagulation) factor 9 cDNA which codes for 461 amino acid has been cloned by screening human fetal liver cDNA library using PCR. This 1.4 kb cDNA spanning from the ATG initiation codon to the TAA termination codon was cloned into bacterial .expression vector pGEX-2T, generating pGEX-F9 plasmid. The plasmid pGEX-F9 expresses about 73 kDa GST (Glutathione S-transferase)-Factor 9 fusion protein when introduced into E. coli. Western blot analysis using polyclonal antibody raised against human factor 9 confirmed this fusion protein contains factor 9 protein. The level of GST-factor 9 expression was about 20% of total protein and the purification of fusion protein was efficiently achieved by using GST agarose bead based on one step purification protocol.

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Expression Patterns and Isolation of Genomic DNA of a Metallothionein-like Gene from Citrus (Citrus unshiu Marc. cv. Miyagawa) (감귤에서 분리한 Metallothionein 유전자의 발현분석 및 게놈 DNA)

  • 김인중
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.231-237
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    • 2001
  • A cDNA clone encoding metallothionein-like protein (CitMT45), which was reported by Moriguchi et al. (1998), was isolated from Citrus fruits cDNA library through differential screening. Our cDNA clone has longer 5'untranslated region, compared to it isolated by Moriguchi et al. (1998). RNA blot analysis showed that the mRNA was abundant in fleshes than peels, leaves, and flowers, as a single transcript. However, regardless of tissue types, the blots showed the similar expression patterns in the process of development with some different profile. These results suggest that CitMT45 may play important roles in the development and/or senescence of various tissues of Citrus. A genomic clone corresponding to CitMT45 was isolated and found to have three exons and two introns. A primer extension analysis suggested that the transcription of CitMT45 gene was started at three start sites with different degrees. The 5'-flanking region was shown to contain a putative metal regulatory element (MRE) and low- temperature responsive element which suggests the possibility of metal-and cold-regulated transcription, respectively.

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Construction and Analysis of a DNA Microarray for the Screening of Biosynthetic Genes of Secondary-Metabolites formation in Streptomyces (방선균 유래 이차대사 생합성 유전자 분석용 DNA Microarray 제작 및 해석)

  • Nam Soo Jung;Kang Dae-Kyung;Rhee Ki Hyeong;Kim Jong-Hee;Kang Sang Sun;Chang Yong Keun;Hong Soon-Kwang
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.105-111
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    • 2005
  • Streptomyces produces many kinds of secondary-metabolites including antibiotics. Screening of a new compound and elucidation of a biosynthetic pathway for the secondary metabolites are very important fields of biology, however, there is a main problem that most of the identified compounds are already researched compounds. To solve these problems, a microarray system that is based on the data related to the biosynthetic genes for secondary-metabolites was designed. For the main contents of DNA microarray, the important genes for the bio-synthesis of aminoglycosides, polyenes group, enediyne group, alpha-glucosidase inhibitors, glycopeptide group, and orthosomycin group were chosen. A DNA microarray with 69 genes that were involved in the bio-synthesis for the antibiotics mentioned above was prepared. The usability of the DNA microarray was confirmed with the chromosomal DNA and total RNA extracted from S. coelicolor whose genomic sequence had already been reported.

Cloning of hnRNP E1 cDNA via yeast two-hybrid system and a study on protein-protein interaction between hnRNP E1 and hnRNP K (이스트 two-hybrid 시스템을 이용한 hnRNP E1 cDNA의 클로닝과 hnRNP E1-hnRNP K 상호결합에 대한 연구)

  • Choi, Mie-Young
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.9 no.6
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    • pp.1795-1799
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    • 2008
  • The heterogeneous nuclear ribonucleoprotein K (hnRNP K) is a component of hnRNP complexes. This protein binds strongly to cytidine-rich RNA/DNA sequences. It is a nucleocytoplasmic shuttling protein. To investigate the functions of hnRNP K, I searched for hnRNP K-interacting proteins in HeLa cDNA library using a yeast two-hybrid screening system. One of the cDNA clones is identical to human hnRNP E1 (poly(rC) binding protein 1) cDNA (GenBank accession number XM_031585). In this study, hnRNP K is shown to specifically interact with hnRNP E1 in yeast two-hybrid system and in vitro biochemical assay.