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Selection of precore mutants during lamivudine treatment in patients with chronic hepatitis B.
Cheong, Jae Youn,Cho, Sung Won,Yoo, Jun Hwan,Hong, Sun Pyo,Kim, Soo-Ok,Yoo, Wang Don,Kim, Jin Hong G. Thieme 2008 Hepato-gastroenterology Vol.55 No.84
<P>BACKGROUND/AIMS: Evolution of precore genes can occur during lamivudine therapy in HBV infection. This study investigated the changes in precore regions in patients treated with lamivudine and the pattern during relapse. METHODOLOGY: The sequences of codon 28 in precore region in serial samples of 16 patients with HBV (11 HBeAg-positive and 5 HBeAg-negative) treated with lamivudine were analyzed by restriction fragment mass polymorphism. RESULTS: Among 9 patients who had wild-type virus, the wild-type virus was replaced by A1896 during relapse after initial treatment in 2 patients, and a pure population with A1896 selected during relapse in all 4 patients with mixed infection. In 5 patients with A1896 during relapse, 3 patients initially reverted to wild-type and later selected A1896, and 2 patients maintained A1896 during lamivudine retreatment. In 8 patients showing HBeAg negative reactivation, 3 patients showed A1896 and 5 patients showed wildtype virus. CONCLUSIONS: Lamivudine therapy induced initial reversion from precore mutants to wild-type virus, but precore mutants reappeared in patients infected with precore mutants. In some patients infected by wildtype HBV, wild-type HBV was replaced by precore mutants, resulting in a flare-up of hepatitis after cessation of lamivudine administration, and HBeAg negativity did not always correspond to the presence of precore mutants.</P>
대장균에서 발현시킨 사람 파필로마 바이러스 18 형 E6 단백질의 분리정제와 사람 혈청내의 18 형 E6 항체 검색에의 이용
정태화,손우익,박순희,강주현,유왕돈,진승원 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
Human papillomavirus type 16 and 18 encode two major transforming proteins E6 and E7. Recombinant E6 protein for human papillomavirus type 18(HPV 18) E6 gene was expressed in E. coli in large quantity as authentic size, non fused form of protein. However, the protein was produced in inclusion bodies. Therefore this bacterially produced E6 protein was solublized in SDS solution and purified to the homogeneity by gel filtration column chromatographies using Sephacryl S-200 column. The yield of purified 18 E6 recombinant protein was approximately 30 ㎎/ℓ liter culture. This purified recombinant 18E6 protein was used to determine the presence of antibodies that might have been produced against HPV 18 infections in the human sera. We tested and compared sera obtained from both cervical cancer patients and people having no apparent cervical cancers. These were accomplished using Western blot. Antibodies reactive to recombinant HPV 18E6 protein were frequently detected in cases with cervical carcinoma(14 out of 20, 70%), compared to people with no apparent carcinoma(4 out of 9, 44%).
( Sang Hoon Ahn ),( Ji Yong Chun ),( Soo Kyung Shin ),( Jun Yong Park ),( Wang Don Yoo ),( Sun Pyo Hong ),( Soo Ok Kim ),( Kwang Hyub Han ) 대한간학회 2013 Clinical and Molecular Hepatology(대한간학회지) Vol.19 No.4
Background/Aims: Molecular diagnostic methods have enabled the rapid diagnosis of drug-resistant mutations in hepatitis B virus (HBV) and have reduced both unnecessary therapeutic interventions and medical costs. In this study we evaluated the analytical and clinical performances of the HepB Typer-Entecavir kit (GeneMatrix, Korea) in detecting entecavir-resistance-associated mutations. Methods: The HepB Typer-Entecavir kit was evaluated for its limit of detection, interference, cross-reactivity, and precision using HBV reference standards made by diluting high-titer viral stocks in HBV-negative human serum. The performance of the HepB Typer-Entecavir kit for detecting mutations related to entecavir resistance was compared with direct sequencing for 396 clinical samples from 108 patients. Results: Using the reference standards, the detection limit of the HepB Typer-Entecavir kit was found to be as low as 500 copies/mL. No cross-reactivity was observed, and elevated levels of various interfering substances did not adversely affect its analytical performance. The precision test conducted by repetitive analysis of 2,400 replicates with reference standards at various concentrations showed 99.9% agreement (2398/2400). The overall concordance rate between the HepB Typer-Entecavir kit and direct sequencing assays in 396 clinical samples was 99.5%. Conclusions: The HepB Typer-Entecavir kit showed high reliability and precision, and comparable sensitivity and specificity for detecting mutant virus populations in reference and clinical samples in comparison with direct sequencing. Therefore, this assay would be clinically useful in the diagnosis of entecavir-resistance-associated mutations in chronic hepatitis B.
Noh, Kap-Soo,Kim, Jong-Wan,Ha, Suk-Hoon,Yoo, Wang-Don,Jeon, Weong-Joong,Kim, Hyun-Su The Korean Society for Biotechnology and Bioengine 1999 Biotechnology and Bioprocess Engineering Vol.4 No.1
Although many pharmaceutically useful proteins are produced in E. coli expression system, it is very are rare for the system to be used in the production of diagnostic antigen due to a major problem, i.e., false-positive reaction of e. coli host-derived proteins contaminating purified diagnostic antigen with human sera. The N (nucleocapsid) protein of Seoul virus causing haemorrhagic fever with renal syndrome (HFRS) was produced in E. coli BL21 (DE3), and used for the detection of N protein-specific antibodies in human sera. Using the N protein as a diagnostic antigen of HFRS, the false-positive reaction was cleared by merely mixing the test sera with the extract of E. coli host strain not harboring expression plasmid.