• 미생물학회지
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• 제55권1호
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• pp.25-32
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• 2019

Influenza A (H1N1) virus caused a worldwide pandemic in 2009-2010 and still remains in seasonal circulation. Continuous surveillance activities are encouraged in the post pandemic phase to watch over the trend of occurrence every year, this is better to be done by a rapid and sensitive method for its detection. This study was conducted to detect proportions of occurrence of influenza A virus (H1N1) in patients with influenza-like illness. Samples from 500 patients with influenza or influenza-like clinical presentation were tested by real-time reverse transcription polymerase chain reaction (RT-PCR) and virus tissue culture. Among the total 500 participants, 193 (38.6%) were females and 307 (61.4%) males. Seventy-one patients (14.2%) were positive for H1N1 virus infection with real-time RT-PCR while 52 (10.4%) were positive by tissue culture. Non-statistically significant relation was found between age and gender with the positivity of H1N1. Sensitivity and specificity of real-time RT-PCR was 98.08% and 95.54%, respectively, in comparison to virus isolation with accuracy 95.8%. This study showed that H1N1 virus was responsible for a good proportion of influenza during the post-pandemic period. Real-time RT-PCR provides rapidity and sensitivity for the detection of influenza A virus (H1N1) compared with virus isolation and thus it is recommended as a diagnostic tool.

• Journal of Microbiology and Biotechnology
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• 제28권3호
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• pp.465-472
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• 2018

Our aim was to determine the detection rate of respiratory viruses (RVs) in feces of patients with acute viral respiratory infection (AVRI) and the detection rate of diarrheal viruses (DVs) in nasopharyngeal samples from patients with acute viral gastroenteritis. The relationships between the presence of fecal RVs or nasopharyngeal DVs and their impacts on the clinical severity were also investigated. A total of 144 fecal specimens were collected from AVRI patients and 95 nasopharyngeal specimens were collected from acute viral gastroenteritis patients. Clinical characteristics and laboratory profiles were compared between subgroups on the basis of the presence or absence of virus in the specimens. The detection rate of RVs in feces was 17.4% (25/144), whereas the detection rate for viruses identical to the respiratory pathogen was 10.4% (identical group, 15/144). Within the identical group, adenovirus (86.7%, 13/15) was most commonly found. Patients in the identical group showed statistically higher values for C-reactive protein, mean age, increased frequency of vomiting, and decreased frequency of chest film involvement and cough (p < 0.05). The detection rate of nasopharyngeal DVs among acute viral gastroenteritis patients was 19.0% (18/95), and in the identical group it was 15.8% (15/95). Norovirus group II and enteric adenovirus were the major pathogens detected in the identical group. There were no significant differences in clinical characteristics and laboratory profiles between the subgroups. In conclusion, the major pathogens of fecal RV and nasopharyngeal DV were adenovirus and norovirus group II, respectively. However, their relationship with the clinical symptoms or disease severity is unclear.

• 한국식물병리학회지
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• 제14권6호
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• pp.636-641
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• 1998

Digoxigenin (DIG) was used to prepare nucleic acid probe for the detection of RNA of potato leafroll virus (PLRV) in the potato leaf extracts. The 0.6 kb coat protein (CP) gene cDNA of PLRV in plasmid pSPT 18 vector was labeled with digoxigenin by in vitro run-off transcription and then used for cRNA probe. In the several buffers tested for increase the total RNA extraction efficiency AMES buffer was the most suitable for this detection method. The RNA extracts from potato leaves shown symptoms of PLRV were dot blotted onto nylon membrane and hybridized with labeled RNA probes. After hybridization, labeled RNA bound to PLRV RNA on membrane was detected with anti-digoxigenin alkaline phosphatase. 5-bromo-4-chloro-3-indolyl-phosphate/nitroblue tetrazolium (NBT) salt and CSPD were used as substrate for colorimetric and film exposure detection, respectively. These detection methods were very sensitive allowing for detection of 1/32 diluted total RNA extract from 100 mg leaf tissue.

• The Plant Pathology Journal
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• 제22권2호
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• pp.168-173
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• 2006

To develop the diagnostic method for the viral infection in apple, the partial genes corresponding to the N-terminal region of RNA polymerase of Apple stem grooving virus (ASGV) and coat protein of Apple chlorotic leaf spot virus (ACLSV) were characterized from the infected apple cultivars in Korea. Based on the nucleotide sequences of the characterized partial genes, the virus gene-specific primers were designed for the detection of ASGV and ACLSV infected in species of Malus. The RT-PCR using the primers for the genes of ASGV and ACLSV successfully gave rise to 404 and 566 bp DNA fragments, respectively. Using those viral gene-specific primers, the multiplex RT-PCR assays were also established to diagnose the mixed infection by ASGV and ACLSV simultaneously. Furthermore, the control primers, which have to be included for the RT-PCR as an internal control, were designed using the nucleotide sequence of the gene encoding elongation factor $1{\alpha}(EF1{\alpha})$. This multiplex RT-PCR including the control primers provides more reliable, rapid and sensitive assay for the detection of ASGV and ACLSV infected in Korean apple cultivars.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2014년도 제46회 동계 정기학술대회 초록집
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• pp.419-419
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• 2014

Interdigitated nanogap device (IND) is an attractive tool for biomolecular detection due to its huge on-off signal ratio, great tolerance to the variation in biochemical environment, and relatively simple implementation processes. Here, we report on the IND-based detection of Influneza A virus by sandwich immunoassay. The INEs were fabricated by photo lithography followed by the in-house chemical lithographic technique for the narrowing the initial gap distance. The surface of the silicon oxide between the two gold electrodes was chemically modified to immobilize primary antibodies for the immuno-specific interaction with the influenza A virus antigen. After immersing the functionalized-IND into the sample solution containing the influenza A virus, the device was exposed to the secondary antibody conjugated Au nanoparticles (Au NPs). The INDs showed a huge jump in the electric conductance when the sample solution contained the influenza A virus of the concentration as low as 10 ng/mL. We hope that this IND-based sensing can be applied to the development of simple and reliable diagnostic means of influenza viruses.

• Bulletin of the Korean Chemical Society
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• 제35권4호
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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.

• 한국정보처리학회:학술대회논문집
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• 한국정보처리학회 2002년도 춘계학술발표논문집 (하)
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• pp.935-938
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• 2002

이 논문에서는 리눅스 기반 VDPM(Virus Detection Protection Management)시스템을 제안하고 개발한 응용 SW 로 감지, 차단 및 관리 방법을 제시한다. 개발된 VDPM 시스템은 신종 바이러스까지 탐지하는 모든 종류의 바이러스 탐지(VDPM_hawkeye) 모듈, Virus 첵크하는 감시 및 Virus 첵크후 교정, 제거하는 방지(VDPM_medic)모듈, DB를 update하는 기능을 가지는 관리(VDPM_manager)모듈과 원격 DB 관리 및 Virus 결과 보고 기능(VDPM_reporter) 모듈로 되어 있으며 지능적인 Virus 방지 시스템을 구현 하였다.

• Pediatric Infection and Vaccine
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• 제21권1호
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• pp.22-28
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• 2014

목적: 임상적으로 중증 세균성 감염과 바이러스성 질환의 감별이 어려운 어린 영아에서 호흡기 바이러스를 검출하고 이것과 연관된 임상적 위험인자를 분석하였다. 방법: 2011년 9월부터 2012년 8월까지 생후 90일 이하 영아 중 패혈증을 포함한 감염성 질환이 의심된 227명을 대상으로 비인두 검체를 채취하였으며 임상적 특성에 대한 후향적 연구를 시행하였다. 채취한 검체 내 호흡기 바이러스의 검출은 real-time PCR 검사를 통해 측정되었다. 결과: 총 157명(69.2%)의 영아에서 한 가지 이상의 호흡기 바이러스가 검출되었다. 빈도는 RSV (75명), RV (42명), influenza virus (18명), parainfluenza virus (15명), human metapneumovirus (9명), coronavirus (9명), adenovirus (4명), bocavirus (3명) 순이었다. 이 중 24명(10.6%)에서 세균성 감염을 진단하였다. 기침, 호흡기 질환의 가족력이 있는 경우 혹은 가을/겨울 철에 호흡기 바이러스가 의미있게 높은 빈도로 검출되었으며 로지스틱 회귀분석에서도 같은 경향을 확인하였다. 가을과 겨울에는 세균성 감염 환자보다 그렇지 않은 환자에서 호흡기 바이러스 검출이 유의하게 많은 것을 알 수 있었다. 결론: 호흡기 바이러스는 감염성 질환이 의심되어 입원한 어린 영아의 중요한 병원체이며 그 검출률은 호흡기 증상, 가을/겨울철 발생, 호흡기 증상의 가족력이 있는 환자에서 유의하게 높았다.

• Journal of Microbiology and Biotechnology
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• 제22권8호
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• pp.1165-1169
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• 2012

In April 2009, the H1N1 pandemic influenza virus emerged as a novel influenza virus. The aim of this study was to compare the performances of several molecular assays, including conventional reverse transcription polymerase chain reaction (RT-PCR), two real-time reverse transcription (rRT)-PCRs, and two multiplex RTPCRs. A total of 381 clinical specimens were collected from patients (223 men and 158 women), and both the Seeplex RV7 assay and rRT-PCR were ordered on different specimens within one week after collection. The concordance rate for the two methods was 87% (332/381), and the discrepancy rate was 13% (49/381). The positive rates for the molecular assays studied included 93.1% for the multiplex Seeplex RV7 assay, 93.1% for conventional reverse transcription (cRT)-PCR, 89.7% for the multiplex Seeplex Flu ACE Subtyping assay, 82.8% for protocol B rRT-PCR, and 58.6% for protocol A rRT-PCR. Our results showed that the multiplex Seeplex assays and the cRT-PCR yielded higher detection rates than rRT-PCRs for detecting the influenza A (H1N1) virus. Although the multiplex Seeplex assays had the advantage of simultaneous detection of several viruses, they were time-consuming and troublesome. Our results show that, although rRT-PCR had the advantage, the detection rates of the molecular assays varied depending upon the source of the influenza A (H1N1)v virus. Our findings also suggest that rRT-PCR sometimes detected virus in extremely low abundance and thus required validation of analytical performance and clinical correlation.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제16권3호
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• pp.162-170
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• 2013

Purpose: To detect major acute gastroenteritis virus (rotavirus, norovirus, astrovirus, and enteric adenovirus) and non-enteric type of adenovirus (AdV) in the stools of intussusception patients and to investigate the clinical role of detected viruses. Methods: From March 2012 to February 2013, major acute gastroenteritis virus and non-enteric type of AdV were isolated from stool samples that collected from 44 patients treated for intussusception in Chungnam National University Hospital. Patients were divided according to age and isolated virus. Results: Virus was detected in 28 (63%) stool specimens. The virus detection rate was significantly lower in patients aged under 12 months (p = 0.04). Twenty-two patients (78.6%) had non-enteric adenovirus, 4 (14.3%) had norovirus, 1 (3.6%) had sapovirus, and 1 (3.6%) had astrovirus. AdV subgroup C (AdV 1, 2, 5, and 6) comprised the majority with 20 cases (90.9%). A monthly increment-and-decrement pattern of intussusception was similar to that of viral detection in the stool samples. Enema reductions were successful in 39 patients and surgical manual reductions were performed in 5 patients. Virus was detected in 24 patients (61.5%) of enema reduction group and 4 patients (80.0%) of surgical manual reduction group. All of the detected viruses were non-enteric adenovirus subgroup C (AdV 1, 5, and 6) in surgical reduction patients. Conclusions: The virus detection rate was high in the stools of intussusception patients. The pattern of seasonal intussusception occurrence rate was parallel with seasonal these viral detection rate in the stool samples. These findings suggest that viral infection plays an important role in the development of intussusception and further research is warranted.