• Title/Summary/Keyword: virus validation

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Reliability and Validity of COVID-19 Stress Scale in Korean Nursing Students (한국어판 간호대학생의 COVID-19 스트레스 측정도구 신뢰도 및 타당도 검증)

  • Ham, Mi-Young;Lee, So Jung;Lim, So-Hee
    • Journal of the Korean Society of School Health
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    • v.35 no.1
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    • pp.1-10
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    • 2022
  • Purpose: This study investigated the validity and reliability of the Korean version of COVID-19 Stress Scale (K-CSS) in nursing students in Korea. Methods: The subjects of the study were 319 nursing students from three universities located in the metropolitan area. Data were collected for two months using self-reporting questionnaires. The study verified the scale's content validity, construct validity, concurrent validity and reliability, using SPSS 20.0 and AMOS 20.0. Results: K-CSS was comprised of seven factors, with a total of 25 questions; six about socio-economic consequences, four about Xenophobia, five about compulsive checking, three about traumatic stress, three about contamination, two about danger of the virus, and two about danger regarding the healthcare system. K-CSS was validated with confirmatory factor analysis (x2/df=2.32, CFI=.94, GFI=.87, NFI=.90, RMR=.07, RMSEA=.06, TLI=.93). Furthermore, the reliability verification showed a Cronbach's α of 0.87, confirming that the Korean version of the tool was very reliable. Conclusion: This study shows that K-CSS is a valid and reliable instrument to assess nursing students in Korea.

A bioinformatic approach to identify pathogenic variants for Stevens-Johnson syndrome

  • Muhammad Ma'ruf;Justitia Cahyani Fadli;Muhammad Reza Mahendra;Lalu Muhammad Irham;Nanik Sulistyani;Wirawan Adikusuma;Rockie Chong;Abdi Wira Septama
    • Genomics & Informatics
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    • v.21 no.2
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    • pp.26.1-26.9
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    • 2023
  • Stevens-Johnson syndrome (SJS) produces a severe hypersensitivity reaction caused by Herpes simplex virus or mycoplasma infection, vaccination, systemic disease, or other agents. Several studies have investigated the genetic susceptibility involved in SJS. To provide further genetic insights into the pathogenesis of SJS, this study prioritized high-impact, SJS-associated pathogenic variants through integrating bioinformatic and population genetic data. First, we identified SJS-associated single nucleotide polymorphisms from the genome-wide association studies catalog, followed by genome annotation with HaploReg and variant validation with Ensembl. Subsequently, expression quantitative trait locus (eQTL) from GTEx identified human genetic variants with differential gene expression across human tissues. Our results indicate that two variants, namely rs2074494 and rs5010528, which are encoded by the HLA-C (human leukocyte antigen C) gene, were found to be differentially expressed in skin. The allele frequencies for rs2074494 and rs5010528 also appear to significantly differ across continents. We highlight the utility of these population-specific HLA-C genetic variants for genetic association studies, and aid in early prognosis and disease treatment of SJS.

Determination of N-nitrosodimethylamine in zidovudine using high performance liquid chromatography-tandem mass spectrometry

  • Yujin Lim;Aelim Kim;Yong-Moon Lee;Hwangeui Cho
    • Analytical Science and Technology
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    • v.36 no.6
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    • pp.281-290
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    • 2023
  • Zidovudine is an antiretroviral agent prescribed for the prevention and treatment of human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS). It is typically recommended to be used in combination with other antiretroviral drugs. Zidovudine has the potential to generate N-nitrosodimethylamine (NDMA) in the presence of dimethylamine and nitrite salt under acidic reaction conditions during the drug manufacturing process. NDMA is a potent human carcinogen that may be detected in drug substances or drug products. An analytical method was developed to determine NDMA in pharmaceuticals including zidovudine using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The analysis involved reversed-phase chromatography on a Kinetex F5 column with a mobile phase comprising water-acetonitrile mixtures. The detection of positively charged ions was conducted using atmospheric pressure chemical ionization (APCI). The calibration curve demonstrated excellent linearity (r = 0.9997) across the range of 1-50 ng/mL with a highly sensitive limit of detection (LOD) at 0.3 ng/mL. The developed method underwent thorough validation for specificity, linearity, accuracy, precision, robustness, and system suitability. This sensitive and specific analytical method was applied for detecting NDMA in zidovudine drug substance and its formulation currently available in the market, indicating its suitability for drug quality management purposes.

Development of sequential sampling plan for Frankliniella occidentalis in greenhouse pepper (고추 온실에서 꽃노랑총채벌레의 축차표본조사법 개발)

  • SoEun Eom;Taechul Park;Kimoon Son;Jung-Joon Park
    • Korean Journal of Environmental Biology
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    • v.40 no.2
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    • pp.164-171
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    • 2022
  • Frankliniella occidentalis is an invasive pest insect, which affects over 500 different species of host plants and transmits viruses (tomato spotted wilt virus; TSWV). Despite their efficiency in controling insect pests, pesticides are limited by residence, cost and environmental burden. Therefore, a fixed-precision level sampling plan was developed. The sampling method for F. occidentalis adults in pepper greenhouses consists of spatial distribution analysis, sampling stop line, and control decision making. For sampling, the plant was divided into the upper part(180 cm above ground), middle part (120-160 cm above ground), and lower part (70-110 cm above ground). Through ANCOVA, the P values of intercept and slope were estimated to be 0.94 and 0.87, respectively, which meant there were no significant differences between values of all the levels of the pepper plant. In spatial distribution analysis, the coefficients were derived from Taylor's power law (TPL) at pooling data of each level in the plant, based on the 3-flowers sampling unit. F. occidentalis adults showed aggregated distribution in greenhouse peppers. TPL coefficients were used to develop a fixed-precision sampling stop line. For control decision making, the pre-referred action thresholds were set at 3 and 18. With two action thresholds, Nmax values were calculated at 97 and 1149, respectively. Using the Resampling Validation for Sampling Program (RVSP) and the results gained from the greenhouses, the simulated validation of our sampling method showed a reasonable level of precision.

Real-Time PCR for Quantitative Detection of Bovine Herpesvirus Type 1 (Bovine Herpesvirus Type 1 정량 검출을 위한 Real-Time PCR)

  • Lee, Dong-Hyuck;Jeong, Hyo-Sun;Lee, Jung-Hee;Kim, Tae-Eun;Lee, Jung-Suk;Kim, In-Seop
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.14-21
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    • 2008
  • Bovine blood, cell, tissue, and organ are used as raw materials for manufacturing biopharmaceuticals, tissue engineered products, and cell therapy. Manufacturing processes for the biologicals using bovine materials have the risk of viral contamination. Therefore viral validation is, essential in ensuring the safety of the products. Bovine herpesvirus type 1 (BHV-1) is the most common bovine pathogen found in bovine blood, cell, tissue, and organ. In order to establish the validation system for the BHV-1 safety of the products, a real-time PCR method was developed for quantitative detection of BHV-1 in raw materials, manufacturing processes, and final products as well as BHV-1 clearance validation. Specific primers for amplification of BHV-1 DNA was selected, and BHV-1 DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $2\;TCID_{50}/ml$. The real-time PCR method was validated to be reproducible and very specific to BHV-1. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with BHV-1. BHV-1 DNA could be quantified in CHO cell as well as culture supernatant. Also the real-time PCR assay could detect $10\;TCID_{50}/ml$ of BHV-1 artificially contaminated in bovine collagen. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of BHV-1 contamination during the manufacture of biologics.

Loop-Mediated Isothermal Amplification Targeting Actin DNA of Trichomonas vaginalis

  • Goo, Youn-Kyoung;Shin, Won-Sik;Yang, Hye-Won;Joo, So-Young;Song, Su-Min;Ryu, Jae-Sook;Kong, Hyun-Hee;Lee, Won-Ki;Chung, Dong-Il;Hong, Yeonchul
    • Parasites, Hosts and Diseases
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    • v.54 no.3
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    • pp.329-334
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    • 2016
  • Trichomoniasis caused by Trichomonas vaginalis is a common sexually transmitted disease. Its association with several health problems, including preterm birth, pelvic inflammatory disease, cervical cancer, and transmission of human immunodeficiency virus, emphasizes the importance of improved access to early and accurate detection of T. vaginalis. In this study, a rapid and efficient loop-mediated isothermal amplification-based method for the detection of T. vaginalis was developed and validated, using vaginal swab specimens from subjects suspected to have trichomoniasis. The LAMP assay targeting the actin gene was highly sensitive with detection limits of 1 trichomonad and 1 pg of T. vaginalis DNA per reaction, and specifically amplified the target gene only from T. vaginalis. Validation of this assay showed that it had the highest sensitivity and better agreement with PCR (used as the gold standard) compared to microscopy and multiplex PCR. This study showed that the LAMP assay, targeting the actin gene, could be used to diagnose early infections of T. vaginalis. Thus, we have provided an alternative molecular diagnostic tool and a point-of-care test that may help to prevent trichomoniasis transmission and associated complications.

Molecular Application in Psychiatry (정신과의 분자생물학 적용)

  • Choi, Ihn-Geun
    • Korean Journal of Biological Psychiatry
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    • v.7 no.2
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    • pp.115-122
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    • 2000
  • The development of molecular biology has brought many changes in psychiatry. Molecular biology makes us possible to know the cause of mental disorders that provide the way to prevent the disorders, and to develop various accurate diagnostic and treatment methods for mental disorders. The author discusses the concept, cause, and treatment of mental disorders in the aspect of molecular biology. Importing the methods of molecular biology into psychiatry, we can anticipate to get a number of the goals of psychiatric genetics, including identification of specific susceptibility genes, clarification of the pathophysiological processes whereby these genes lead to symptoms, establishment of epigenetic factors that interact with these genes to produce disease, validation of nosological boundaries that more closely reflect the actions of these genes, and development of effective preventive and therapeutic interventions based on genetic counseling, gene therapy, and modification of permissive or protective environmental influences. In addition to their capacity to accelerate the discovery of new molecules participating in the nervous system's response to disease or to self-administered drugs, molecular biological strategies can also be used to determine how critical a particular gene product may be in mediating a cellular event with behavioral importance. Molecular biology probably enables us discover the environmental factors of mental disorders and allow rational drug design and gene therapies for mental disorders, by isolation of gene products that facilitate a basic understanding of the pathogenesis of these disorders. A specific genetic linkage may suggest a novel class of drugs that has not yet been tried. With respect to gene therapy, the hypothetical method would use a gene delivery system, most likely a modified virus, to insert a functional copy of a mutant gene into those brain cells that require the gene for normal function.

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Molecular and Circulatory Expression of Insulin Growth Factors in Indian Females with Advanced Cervical Cancer

  • Sharma, Manoj;Satyam, Abhigyan;Abhishek, Ashu;Khan, Rehan;Rajappa, Medha;Sharma, Alpana
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6475-6479
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    • 2012
  • Background: Recent studies have demonstrated an association between insulin growth factor (IGF) and insulin growth factor binding protein-3 (IGFBP-III) serum levels and increased risk for various cancers. However, little information is available on clinical implications of the IGF system in Indian patients with cervical cancer. This study explored associations by analyzing their expression profiles in cervical cancer cases. Materials and Methods: Totals of 50 patients with advanced cervical cancer and 40 healthy controls were enrolled. Human papillomavirus (HPV) and cervical biopsy sample were obtained from all participating women. Circulatory levels were estimated by ELISA and the tissue expression was assessed using RT-PCR and Western blotting. Results: Levels of IGF-I and II showed significant increase whereas IGFBP-III showed significant decline in all patients as compared to controls. Spearman correlation analysis between IGFs and HPV status showed significant correlations. Conclusions: We demonstrated elevated circulating levels and tissue expression of IGF-I and IGF-II in advancer cancer cervix patients, as compared with controls, with a converse trend being apparent for IGFBP-III. In future, associations of the IGF system and clinical outcome of cervical cancer patients in post treatment samples might point to significance in disease mapping as a prognostic marker after validation with a larger patient series.

Validation of One-Step Real-Time RT-PCR Assay in Combination with Automated RNA Extraction for Rapid Detection and Quantitation of Hepatitis C Virus RNA for Routine Testing in Clinical Specimens

  • KIM BYOUNG-GUK;JEONG HYE-SUNG;BAEK SUN-YOUNG;SHIN JIN-HO;KIM JAE-OK;MIN KYUNG-IL;RYU SEUNG-REL;MIN BOK-SOON;KIM DO-KEUN;JEONG YONG-SEOK;PARK SUE-NIE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.595-602
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    • 2005
  • A one-step real-time quantitative RT-PCR assay in combination with automated RNA extraction was evaluated for routine testing of HCV RNA in the laboratory. Specific primers and probes were developed to detect 302 bp on 5'-UTR of HCV RNA. The assay was able to quantitate a dynamic linear range of $10^7-10^1$ HCV RNA copies/reaction ($R^2=0.997$). The synthetic HCV RNA standard of $1.84{\pm}0.1\;(mean{\pm}SD)$ copies developed in this study corresponded to 1 international unit (IU) of WHO International Standard for HCV RNA (96/790 I). The detection limit of the assay was 3 RNA copies/reaction (81 IU/ml) in plasma samples. The assay was comparable to the Amplicor HCV Monitor (Monitor) assay with correlation coefficient r=0.985, but was more sensitive than the Monitor assay. The assay could be completed within 3 h from RNA extraction to detection and data analysis for up to 32 samples. It allowed rapid RNA extraction, detection, and quantitation of HCV RNA in plasma samples. The method provided sufficient sensitivity and reproducibility and proved to be fast and labor-saving, so that it was suitable for high throughput HCV RNA test.

Phenotypic and Genotypic Detection of Metallo-β-Lactamase Producing Pseudomonas aeruginosa

  • Yang, Byoung-Seon;Hong, Keun-Seok;Jung, Seung-Bong;Kwon, Young-Hoon;Jeong, Jong-Yoon;Lee, Min-Joo;Lee, Hye-In;Park, Mi-Seon;Choi, Seung-Gu
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.2
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    • pp.81-85
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    • 2012
  • This study was undertaken to evaluate phenotypic and genotypic methods for detection of Metallo-Beta-Lactamases (MBLs) among nosocomial Pseudomonas aeruginosa. Of the 50 P. aeruginosa isolates from clinical specimens, 20 were evaluated for carbapenem resistance and screened for MBL by double-disk synergy test and combined-disk test. Nineteen strains (95%) were found to be MBL producers among the 20 P. aeruginosa. MBL positives were further confirmed by Polymerase Chain Reaction (PCR). For the IMP and VIM types of MBLs, PCR analysis was performed on 19 of the 20, and 10 were positive for VIM MBL type. This study reports the validation of a simple and accurate MBL detection method that can be easily incorporated into the daily routine of a clinical laboratory. Early detection of MBL-carrying organisms, including those with susceptibility to carbapenems, is of paramount clinical importance, as it allows rapid initiation of strict infection control practices as well as therapeutic guidance for confirmed infection.Key Words : Hepatitis A virus (HAV), Anti-HAV, Hospital workers, Prevalence, Vaccination

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