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- 검색키워드
- 저자 : Hyeyoung Wang (검색결과 28 건)
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Wang, Hye-Young,Kim, Hyunjung,Kim, Yeun,Bang, Hyeeun,Kim, Jong-Pill,Hwang, Joo Hwan,Cho, Sang-Nae,Kim, Tae Ue,Lee, Hyeyoung Microbiological Society of Korea 2015 The journal of microbiology Vol.53 No.10
<P>Drug resistance in Mycobacterium leprae is a significant problem in countries where leprosy is endemic. A sensitive, specific, and high-throughput reverse blot hybridization assay (REBA) for the detection of genotypic resistance to rifampicin (RIF) was designed and evaluated. It has been shown that resistance to RIF in M. leprae involves mutations in the rpoB gene encoding the -subunit of the RNA polymerase. The PCR-REBA simultaneously detects both 6 wild-type regions and 5 different mutations (507AGC, 513GTG, 516TAT, 531ATG, and 531TTC) including the most prevalent mutations at positions 507 and 531. Thirty-one clinical isolates provided by Korea Institute of Hansen-s Disease were analyzed by PCR-REBA with RIF resistance of rpoB gene. As a result, missense mutations at codons 507 AGC and 531ATG with 2-nucleotide substitutions were found in one sample, and a missense mutation at codon 516 TAT and ??WT6 (deletion of 530-534) was found in another sample. These cases were confirmed by DNA sequence analysis. This rapid, simple, and highly sensitive assay provides a practical alternative to sequencing for genotypic evaluation of RIF resistance in M. leprae.</P>
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Wang, Hye-Young,Park, Sunyoung,Kim, Sunghyun,Lee, Dongsup,Kim, Geehyuk,Kim, Yeun,Park, Kwang Hwa,Lee, Hyeyoung American Society for Clinical Pathology 2015 American journal of clinical pathology Vol.143 No.3
<P>Objectives: Human papillomavirus (HPV) is a major cause of cervical cancer, which is the second most common cancer in women. HPV E6 initiates degradation of cellular tumor suppressor protein p53, induces human telomerase reverse transcriptase (hTERT) activity, and then leads to progressive cervical carcinogenesis. Methods: In this study, the CervicGen HPV RT-qDX assay (Optipharm, Osong, Republic of Korea), which detects 16 HPV high-risk subtypes (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, and 69), and the CervicGen hTERT RT-qDX assay (Optipharm) were evaluated using 545 ThinPrep (Hologic, Bedford, MO Papanicolaou samples. Results: The positivity for the HPV E6/E7 messenger RNA (mRNA) assay was 94.4%, 95.2%, 82.4%, 46.5%, 25.0%, and 1.1% in squamous cell carcinomas, high-grade squamous intraepithelial lesions (HSILs), atypical squamous cells-cannot exclude HSIL, low-grade squamous intraepithelial lesions, atypical squamous cells of undetermined significance, and normal cytology samples, respectively. Five cervical intraepithelial neoplasia grade 2+ samples were not detected by the HPV E6/E7 mRNA assay, but they exhibited positive signals in the hTERT mRNA assay. Notably, the hTERT mRNA expression level was increased in high-grade cervical lesions but was very low in all 288 normal samples. Conclusions: These data suggest that the combination of HPV E6/E7 and hTERT mRNA expression levels could be used in a complementary manner in diagnosing high-grade cervical lesions and malignant tumors and might be useful as a predictive marker in monitoring low-grade cervical lesions.</P>
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Wang, Hye-young,Kim, Sunghyun,Kim, Jungho,Park, Soon-Deok,Uh, Young,Lee, Hyeyoung American Society for Microbiology 2014 Journal of clinical microbiology Vol.52 No.6
<P>Methicillin-resistant <I>Staphylococcus aureus</I> (MRSA) is the most prevalent cause of bloodstream infections (BSIs) and is recognized as a major nosocomial pathogen. This study aimed to evaluate a newly designed multiplex real-time PCR assay capable of the simultaneous detection of <I>mecA</I>, <I>S. aureus</I>, and coagulase-negative staphylococci (CoNS) in blood culture specimens. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays (M&D, Republic of Korea) use the TaqMan probes 16S rRNA for <I>Staphylococcus</I> spp., the <I>nuc</I> gene for <I>S. aureus</I>, and the <I>mecA</I> gene for methicillin resistance. The detection limit of the multiplex real-time PCR assay was 10<SUP>3</SUP> CFU/ml per PCR for each gene target. The multiplex real-time PCR assay was evaluated using 118 clinical isolates from various specimen types and a total of 350 positive blood cultures from a continuous monitoring blood culture system. The results obtained with the multiplex real-time PCR assay for the three targets were in agreement with those of conventional identification and susceptibility testing methods except for one organism. Of 350 positive bottle cultures, the sensitivities of the multiplex real-time PCR kit were 100% (166/166 cultures), 97.2% (35/36 cultures), and 99.2% (117/118 cultures) for the 16S rRNA, <I>nuc</I>, and <I>mecA</I> genes, respectively, and the specificities for all three targets were 100%. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays are very useful for the rapid accurate diagnosis of staphylococcal BSIs. In addition, the Real-MRSA and Real-MRCoNS multiplex real-time PCR assays could have an important impact on the choice of appropriate antimicrobial therapy, based on detection of the <I>mecA</I> gene.</P>
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Wang, Hye-young,Lee, Dongsup,Park, Sunyoung,Kim, Geehyuk,Kim, Sunghyun,Han, Lin,Yubo, Ren,Li, Yingxue,Park, Kwang Hwa,Lee, Hyeyoung Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.17
Background: Human papillomavirus (HPV) is the most common sexually transmitted infection worldwide and it is responsible for most cases of cervical uterine cancer. Although HPV infections of the cervix do not always progress to cancer, 90% of cervical cancer cases have been found to be associated with high risk HPV (HR-HPV) infection. HPV DNA testing is widely used, along with Papanicolaou (Pap) testing, to screen for cervical abnormalities. However, there are no data on the prevalence of genotype-specific HPV infections assessed by measuring HPV E6/E7 mRNA in women representative of the Chinese population across a broad age range. Materials and Methods: In the present study, we compared the results with the CervicGen HPV RT-qDx assay, which detects 16 HR-HPV genotypes (Alpha-9: HPV 16, 31, 33, 35, 52, and 58; Alpha-7: HPV 18, 39, 45, 51, 59, and 68; and Alpha-5, 6: HPV 53, 56, 66, and 69), and the REBA HPV-ID assay, which detects 32 HPV genotypes based on the reverse blot hybridization assay (REBA) for the detection of oncogenic HPV infection according to cytological diagnosis. We also investigated the prevalence and genotype distribution of HPV infection with a total of 324 liquid-based cytology samples collected in western Shandong province, East China. Results: The overall HPV prevalences determined by HPV DNA and HPV E6/E7 mRNA assays in this study were 79.9% (259/324) and 55.6% (180/324), respectively. Although the positivity of HPV E6/E7 mRNA expression was significantly lower than HPV DNA positivity, the HPV E6/E7 mRNA assay showed greater specificity than the HPV DNA assay (88.6% vs. 48.1%) in normal cytology samples. The prevalence of Alpha-9 (HPV 16, 31, 33, 35, 52, and 58) HPV infection among these women accounted for up to 80.3% and 76.1% of the high-grade lesions detected in the HPV mRNA and DNA tests, respectively. The HR-HPV genotype distribution, based on HPV DNA and E6/E7 mRNA expression by age group in patients with cytologically confirmed lesions, was highest in women aged 40 to 49 years (35.9% for cytologically confirmed cases, Pearson correlation r value=0.993, p<0.001) for high-grade lesions. Among the oncogenic HR-HPV genotypes for all age groups, there was little difference in the distribution of HPV genotypes between the HPV DNA (HPV -16, 53, 18, 58, and 33) and HPV E6/E7 mRNA (HPV -16, 53, 33, 58, and 18) assays. HPV 16 was the most common HPV genotype among women with high-grade lesions. Conclusions: Our results suggest that the HPV E6/E7 mRNA assay can be a sensitive and specific tool for the screening and investigation of cervical cancer. Furthermore, it may provide useful information regarding the necessity for early cervical cancer screenings and the development of additional effective HPV vaccines, such as one for HPV 53 and 58. Additionally, gaining knowledge of HPV distribution may also inform us about ecological changes in HPV after the vaccination.
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Kim, Jungho,Wang, Hye-young,Kim, Seoyong,Park, Soon Deok,Yu, Kwangmin,Kim, Hyo Youl,Uh, Young,Lee, Hyeyoung Elsevier Biomedical 2016 Journal of microbiological methods Vol.128 No.-
<P><B>Abstract</B></P> <P>DNA extraction efficiency affects the success of PCR-based method applications. The Punch-it™ NA-Sample kit for extracting DNA by using paper chromatography is technically easy to use and requires just two reagents and only 10min to complete. The Punch-it™ NA-Sample kit could be offered as a rapid, accurate, and convenient method for extracting bacterial and fungal DNA from blood culture bottles. We compared the efficiencies of the commercial kit (Punch-it™ NA-Sample kit) and an in-house conventional boiling method with Chelex-100 resin for DNA extraction from blood culture bottles. The efficiency of the two DNA extraction methods was assessed by PCR-reverse blot hybridization assay (PCR-REBA, REBA Sepsis-ID) for detecting Gram positive (GP) bacteria, Gram negative (GN) bacteria, and <I>Candida</I> species with 196 positive and 200 negative blood culture bottles. The detection limits of the two DNA extraction methods were 10<SUP>3</SUP> CFU/mL for GP bacteria, 10<SUP>3</SUP> CFU/mL for GN bacteria, and 10<SUP>4</SUP> CFU/mL for <I>Candida</I>. The sensitivity and specificity of the Punch-it™ NA-Sample kit by REBA Sepsis-ID were 95.4% (187/196) and 100% (200/200), respectively. The overall agreement of the two DNA extraction methods was 98.9% (392/396). Three of four samples showing discrepant results between the two extraction methods were more accurately matched up with the Punch-it™ NA-Sample kit based on conventional culture methods. The results indicated that the Punch-it™ NA-Sample kit extracted bacterial and fungal DNA in blood culture bottles and allowed extracted DNA to be used in molecular assay.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Punch-it™ NA-Sample kit and boiling method for DNA extraction were compared. </LI> <LI> Punch-it™ NA-Sample kit extracted bacterial and fungal DNA in blood culture bottles. </LI> <LI> The Punch-it™ NA-Sample kit does not require enzyme to digest cell walls. </LI> </UL> </P>
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Improved Detection of Mycobacterium leprae by One-tube Nested Polymerase Chain Reaction
Hye-Young Wang,Joo-Hwan Whang,Jong-Pill Kim,Jang-Eun Cho,Hyeeun Bang,Hyeyoung Lee,Sang-Nae Cho 대한의생명과학회 2007 Biomedical Science Letters Vol.13 No.4
One-tube nested polymerase chain reaction (PCR) was evaluated for its efficacy in detecting Mycobacterium leprae in biopsy samples from leprosy patients. Primers were derived from the M. leprae-specific element (RLEP) sequences which yield a 230 bp fragment. The specificity and the sensitivity of the one-tube nested PCR were compared with those of single PCR for detecting M. leprae. The results showed that the one-tube nested PCR was about 100 times more sensitive than that of the single indicating the one-tube nested primer sets developed in this study can be an effective screening tool for the detection of M. leprae in clinical diagnostic laboratories.
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Sunyoung Park,Hyeyoung Wang,Sunghyun Kim,Geehyuk Kim,Sungyoung Bong,Hyoungsoon Jang,Sangjung Park,Kooyeon Hwang,Dongsup Lee 대한의생명과학회 2016 Biomedical Science Letters Vol.22 No.3
Investigation of human papillomavirus (HPV) in archival formalin-fixed paraffin-embedded (FFPE) material is important for understanding cervical carcinogenesis. The objective of the present study was to identify the high risk HPVs (HR-HPVs) using HPV E6/E7 mRNA testing from archival tissues in cervical cancer and the relation to HR-HPVs genotypes in paired cervical exfoliated cells. HPV E6/E7 mRNA testing and DNA chip testing were performed in 79 paired cervical FFPE tissues and exfoliated cells from women with histologically confirmed squamous cell carcinoma and adenocarcinoma. Overall agreement in HR-HPVs detection from FFPE samples and cytology samples were 98.5% in HPV 16, 100% in HPV 18, HPV 31, HPV 33, HPV 58, HPV 66, and HPV 68. Type-specific agreement between FFPE samples and cytology samples was 89.1% in HPV positive, 93.5% in HPV 16 and more than 70% in the other HR-HPVs. In conclusion, HR-HPVs were reliably detected in paired FFPE and cytology samples with some variation in type-specific detection.
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PCR-Reverse Blot Hybridization Assay for Screening and Identification of Pathogens in Sepsis
Choi, Yeonim,Wang, Hye-Young,Lee, Gyusang,Park, Soon-Deok,Jeon, Bo-Young,Uh, Young,Kim, Jong Bae,Lee, Hyeyoung American Society for Microbiology 2013 Journal of clinical microbiology Vol.51 No.5
<P>Rapid and accurate identification of the pathogens involved in bloodstream infections is crucial for the prompt initiation of appropriate therapy, as this can decrease morbidity and mortality rates. A PCR-reverse blot hybridization assay for sepsis, the reverse blot hybridization assay (REBA) Sepsis-ID test, was developed; it uses pan-probes to distinguish Gram-positive and -negative bacteria and fungi. In addition, the assay was designed to identify bacteria and fungi using six genus-specific and 13 species-specific probes; it uses additional probes for antibiotic resistance genes, i.e., the <I>mecA</I> gene of methicillin-resistant <I>Staphylococcus aureus</I> (MRSA) and the <I>vanA</I> and <I>vanB</I> genes of vancomycin-resistant enterococci (VRE). The REBA Sepsis-ID test successfully identified clinical isolates and blood culture samples as containing Gram-positive bacteria, Gram-negative bacteria, or fungi. The results matched those obtained with conventional microbiological methods. For the REBA Sepsis-ID test, of the 115 blood culture samples tested, 47 (40.8%) and 49 (42.6%) samples were identified to the species and genus levels, respectively, and the remaining 19 samples (16.5%), which included five Gram-positive rods, were identified as Gram-positive bacteria, Gram-negative bacteria, or fungi. The antibiotic resistances of the MRSA and VRE strains were identified using both conventional microbiological methods and the REBA Sepsis-ID test. In conclusion, the REBA Sepsis-ID test developed for this study is a fast and reliable test for the identification of Gram-positive bacteria, Gram-negative bacteria, fungi, and antibiotic resistance genes (including <I>mecA</I> for MRSA and the <I>vanA</I> and <I>vanB</I> genes for VRE) in bloodstream infections.</P>
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