• Title/Summary/Keyword: Viral load

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Real-time PCR Quantification of White Spot Syndrome Virus (WSSV) and Hepatopancreatic Parvovirus (HPV) Loads in Shrimp and Seawaters of Shrimp Ponds on the West Coast of South Korea

  • Jang, In-Kwon;Gopalakannan, Ayyaru;Suriakala, Kannan;Kim, Jong-Sheek;Kim, Bong-Rae;Cho, Yeong-Rok;Meng, Xian-Hong;Seo, Hyeong-Chul
    • Fisheries and Aquatic Sciences
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    • v.11 no.4
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    • pp.195-204
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    • 2008
  • Viral diseases are major emerging problems of shrimp that have affected the production, and even complete losses for shrimp farms. In this study, we developed a sensitive TaqMan real-time PCR method to quantify white spot syndrome virus (WSSV) and hepatopancreatic parvovirus (HPV) in the shrimp and pond water in which fleshy shrimp, Fenneropenaeus chinensis, and Pacific white shrimp, Litopenaeus vannamei, are reared. WSSV and HPV in pond seawaters ranged from $1.65{\times}10^3$ to $2.43{\times}10^9$ and from 0 to $4.43{\times}10^5$ copies/L of seawater, respectively. Of 20 ponds analyzed, all pond water and shrimp were positive for WSSv. L. vannamei showed higher susceptibility to WSSV than F chinensis. HPV was detected only in the pond water for F chinensis. In shrimp tissue, however, HPV was found in both species, with 23-times higher infection rate in F chinensis than L. vannamei. The total bacterial counts in the pond water ranged from $2.23{\times}l0^3$ to $1.98{\times}l0^5\;CFU/mL$. The variations in total bacterial count for each pond appeared to correlate to the variations of the WSSV load. Statistical analysis indicated that there was no significant difference (P>0.05) between the WSSV load in pond water and shrimp, and there was no relationship between total bacterial load and viral load in the pond water. However, a significant difference (P<0.01) was found between HPV load and L. vannamei and F chinensis pond water.

Development of Diagnostic Techniques for Newcastle Disease in Chickens by In Situ RT-PCR and In Situ Hybridization (In situ RT-PCR 및 In situ hybridization 기법에 의한 닭 뉴캣슬병의 진단법 개발)

  • Park, Nam-Yong;Choi, Hyo-Im;Cho, Ho-Seong;Kang, Sung-Kwi;Cho, Kyoung-Oh;Brown, Corrie
    • Korean Journal of Veterinary Research
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    • v.42 no.3
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    • pp.351-362
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    • 2002
  • Newcastle disease (ND) is a highly contagious infection of poultry, Two pathology-based techniques, in situ RT-PCR and in situ hybridization (ISH) were applied to formalin-fixed, paraffin-embedded tissues from chickens naturally infected with velogenic ND virus (VNDV). Two pairs of primers and a probe for ISH and in situ RT-PCR, respectively, were selected from highly conserved region of matrix gene of NDV. The ISH experiment was carried out using MicroProbe$^{TM}$ capillary action system within 2 hours. In situ RT-PCR was performed using MicroProbe$^{TM}$ capillary action system and GeneAmp In Situ PCR system. With ISH and in situ RT-PCR, viral nucleic acid was detected in the central nervous system of chickens from infected with neurotropic velogenic Newcastle disease virus (NVNDV), whereas viral nucleic acid was detected in various organs or tissues of chickens from infected with viscerotropic velogenic Newcastle disease virus (VVNDV). In the NVND group, positive signals were characteristically defined in the cytoplasm of neuron, vascular endothelial cells, and perivascular mononuclear macrophages in the central nervous system. One of NVND group, chicken from one farm exhibited positive signals in the bronchial epithelium. The VVND group chickens showed positive reaction in the macrophages, vascular endothelium, and bronchiolar epithelium. Markedly, viral nucleic acid was detected in the macrophages of morphologically normal tissues which were peripheral or located in distant areas from lesions. The central nervous system of chickens infected with VVND virus had positive signals in the vascular endothelial cell, perivascular mononuclear macrophages and some neuron. The number and intensity of the positive cells by in situ RT-PCR were more and stronger, respectively, in comparison with those by ISH. Particularly, positive reaction was detected in macrophages infiltrating in cardiac muscle by in situ RT-PCR, but not obtained by ISH. Therefore, these results demonstrated that ISH is a rapid diagnostic method for detection of NDV and in situ RT-PCR can be used as an efficient method for detection of low viral load infection or subclinical viral infection of NDV.

The Detection of ICD p24 Antigen Predicts Bad Prognosis in HIV-1 Infected Patients (인면역결핍바이러스 감염자에서 ICD-p24 항원 탐지가 CD4+T 세포수 및 예후에 미치는 영향)

  • Cho, Young-Keol;Lee, Hee-Jung
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.259-267
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    • 1996
  • In order to evaluate the effect of viral load on the prognosis of human immunodeficiency virus-1 (HIV-1)-infected individuals, immune complex dissociated (ICD) serum p24 antigen (p24) by acid treatment was retrospectively measured for 50 HIV-infected patients for 60 months. Among them, 27 patients were p24 positive (p24+) above 25pg/ml for $40.4{\pm}12$ months and 23 patients were negative (p24-). Follow-up periods from HIV diagnosis were $63.0{\pm}19$ months (range; 40-112) for the p24+ and $68.4{\pm}19$ months (range; 38-106) for the p24-, respectively (P>0.05)Mean CD4+T cell counts in the p24+ group decreased from $473{\pm}$277/ul (median;373) to $157{\pm}150/ul$ (median; 111) for $60{\pm}16$ months (5.3/month P280/ul (median; 476) to $432{\pm}285/ul$ (median;382) for $63{\pm}19$ months (2.5/month, P<0.01). From CD4+T cell count >200/ul, the patient who progressed to AIDS of <200/ul were 13 of 23 (56%) in the p24+ and 4 of 22 (18%) in p24-, respectively (p<0.01). And the number of death in two groups were 6 (22%) and 1 (4%), respectively (p<0.01). Presumed survival in two groups were about 12 and 24.5 years. These data suggest that viral load itself be very important for the prognosis of HIV-infected patients.

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Preparation of Microspheres Encapsulating a Recombinant TIMP-1 Adenovirus and their Inhibition of Proliferation of Hepatocellular Carcinoma Cells

  • Xia, Dong;Yao, Hui;Liu, Qing;Xu, Liang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6363-6368
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    • 2012
  • Objective: The study aim was to prepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) in an adenovirus to investigate its inhibition on the proliferation of hepatocellular carcinoma cells HepG2. Methods: Microspheres were prepared by encapsulating the recombinant TIMP-1 adenovirus into biodegradable PELA. The particle size, viral load, encapsulation efficiency and in-vitro release were measured. Microspheres were used to infect HepG2 cells, then infection efficiency was examined under a fluorescent microscope and ultrastructural changes assessed by TEM. Expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR and proliferation by MTT and cell growth curve assays. Results: We successfully prepared microspheres encapsulating recombinant TIMP-1 adenovirus with a diameter of $1.965{\mu}m$, an encapsulation efficiency of 60.0%, a viral load of $10.5{\times}10^8/mg$ and approximate 60% of virus release within 120 h, the total releasing time of which was longer than 240 h. The microspheres were confirmed to be non-toxic with blank microspheres. Infected HepG2 cells could stably maintain in-vitro expression of TIMP-1, with significantly effects on biological behaviour Conclusion: PELA microspheres encapsulating a recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for polymer/chemistry-based gene therapy of hepatocellular carcinomas.

Negative Correlation between the Prevalence of Norovirus and High Bacterial Loads of Escherichia coli in Oysters Crassostrea gigas

  • Oh, Eun-Gyoung;Song, Ki Cheol;Kim, Sukyung;Park, Kunbawui;Yu, Hongsik
    • Fisheries and Aquatic Sciences
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    • v.18 no.3
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    • pp.235-240
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    • 2015
  • Seasonal variation in the prevalence of norovirus in oysters Crassostrea gigas was investigated and compared to levels of Escherichia coli, a fecal indicator in oysters. Oysters were collected from Iwon-myeon, Taean-gun, Korea, a primary production area for European Union export of oysters between 2013 and 2014. We observed seasonality in the prevalence of norovirus in oysters, with a higher prevalence and viral load detected during winter months. Oysters taken from production areas that complied with the European Union standard for raw consumption (< 230 MPN/100 g of E. coli) had 22.1% of the samples test positive for norovirus (15/68 samples). However, norovirus was not detected in any of the samples (0%, 0/4 samples) that were collected from production sites that exceeded the standard fecal contamination level for raw consumption (> 230 MPN/100 g of E. coli). These results indicated that there is a negative correlation between the prevalence of norovirus and high levels of E. coli in oysters. Therefore, our results suggest that current food safety guidelines using only a bacterial fecal contamination indicator, E. coli, may not adequately assess shellfish production areas for viral and bacterial contamination.

C-Reactive Protein and Respiratory Viral Infection (호흡기 바이러스 감염과 C-Reactive Protein)

  • Jeon, Jae-Sik;Rheem, Insoo;Kim, Jae Kyung
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.1
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    • pp.15-21
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    • 2017
  • C-reactive protein (CRP) levels are not generally associated with viral infections. This study investigated the changes in the CRP level caused by an infection from respiratory virus (RV). Nasopharyngeal samples from hospitalized patients with suspected RV infection were used to measure the CRP levels, virus load, virus-virus co-infection, age, sex, and length of hospital stay (LOS). Abnormal CRP levels were detected in 62.3% (3,608 out of 5,788) of all RV-positive samples. The percentage of patients with abnormal CRP levels tended to increase with age. Furthermore, LOS in patients with abnormal CRP levels was significantly longer than that in patients with normal CRP levels. The frequency of elevated CRP levels differed according to the causative virus and the frequency of abnormal levels increased with age. Moreover, LOS was longer in those with abnormal CRP levels. These data provide important insights into the role of CRP levels in RV infection.

Multifaceted Usage of HPV Related Tests and Products in the Management of Cervical Cancer - a Review

  • Nalliah, Sivalingam;Karikalan, Barani;Kademane, Kumaraswamy
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2145-2150
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    • 2015
  • HPV viruses are integral to the development of cervical cancer. The pathogenesis has been extensively studied. To date, numerous HPV tests and products have been developed and successfully utilized in diagnosis, treatment and prevention of cervical cancer. The HPV DNA test, when combined with other routine cervical cancer screening and diagnostic tests namely exfoliative cytology, visual inspection with acetic acid (VIA) and colposcopy has increased the detection rate of cervical cancer. HPV DNA products could also be measured in other body fluids like urine, lymph node tissue, and serum. HPV association could also be quantified by measuring other parameters like HPV mRNA, viral load, viral integration and methylation status. Vaccination against HPV has been found to decrease the incidence of cervical cancer. Further, therapeutic vaccines for cervical cancer against HPV continue to evolve. All these findings pertaining to HPV could possibly decrease the incidence of cervical cancer in the near future. This review aims to give an overview of the HPV tests and products in use and those under trial currently.

Immune Cells Are Differentially Affected by SARS-CoV-2 Viral Loads in K18-hACE2 Mice

  • Jung Ah Kim;Sung-Hee Kim;Jeong Jin Kim;Hyuna Noh;Su-bin Lee;Haengdueng Jeong;Jiseon Kim;Donghun Jeon;Jung Seon Seo;Dain On;Suhyeon Yoon;Sang Gyu Lee;Youn Woo Lee;Hui Jeong Jang;In Ho Park;Jooyeon Oh;Sang-Hyuk Seok;Yu Jin Lee;Seung-Min Hong;Se-Hee An;Joon-Yong Bae;Jung-ah Choi;Seo Yeon Kim;Young Been Kim;Ji-Yeon Hwang;Hyo-Jung Lee;Hong Bin Kim;Dae Gwin Jeong;Daesub Song;Manki Song;Man-Seong Park;Kang-Seuk Choi;Jun Won Park;Jun-Won Yun;Jeon-Soo Shin;Ho-Young Lee;Ho-Keun Kwon;Jun-Young Seo;Ki Taek Nam;Heon Yung Gee;Je Kyung Seong
    • IMMUNE NETWORK
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    • v.24 no.2
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    • pp.7.1-7.19
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    • 2024
  • Viral load and the duration of viral shedding of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are important determinants of the transmission of coronavirus disease 2019. In this study, we examined the effects of viral doses on the lung and spleen of K18-hACE2 transgenic mice by temporal histological and transcriptional analyses. Approximately, 1×105 plaque-forming units (PFU) of SARS-CoV-2 induced strong host responses in the lungs from 2 days post inoculation (dpi) which did not recover until the mice died, whereas responses to the virus were obvious at 5 days, recovering to the basal state by 14 dpi at 1×102 PFU. Further, flow cytometry showed that number of CD8+ T cells continuously increased in 1×102 PFU-virus-infected lungs from 2 dpi, but not in 1×105 PFU-virus-infected lungs. In spleens, responses to the virus were prominent from 2 dpi, and number of B cells was significantly decreased at 1×105 PFU; however, 1×12 PFU of virus induced very weak responses from 2 dpi which recovered by 10 dpi. Although the defense responses returned to normal and the mice survived, lung histology showed evidence of fibrosis, suggesting sequelae of SARS-CoV-2 infection. Our findings indicate that specific effectors of the immune response in the lung and spleen were either increased or depleted in response to doses of SARS-CoV-2. This study demonstrated that the response of local and systemic immune effectors to a viral infection varies with viral dose, which either exacerbates the severity of the infection or accelerates its elimination.

Viral Load Dynamics After Symptomatic COVID-19 in Children With Underlying Malignancies During the Omicron Wave

  • Ye Ji Kim;Hyun Mi Kang;In Young Yoo;Jae Won Yoo;Seong Koo Kim;Jae Wook Lee;Dong Gun Lee;Nack-Gyun Chung;Yeon-Joon Park;Dae Chul Jeong;Bin Cho
    • Pediatric Infection and Vaccine
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    • v.30 no.2
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    • pp.73-83
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    • 2023
  • Purpose: This study aimed to investigate the viral load dynamics in children with underlying malignancies diagnosed with symptomatic coronavirus disease 2019 (COVID-19). Methods: This was a retrospective longitudinal cohort study of patients <19 years old with underlying hemato-oncologic malignancies that were diagnosed with their first symptomatic severe acute respiratory syndrome coronavirus 2 polymerase chain reaction (PCR)-confirmed COVID-19 infection during March 1 to August 30, 2022. Review of electronic medical records and telephone surveys were undertaken to assess the clinical presentations and transmission route of the patients. Thresholds of negligible likelihood of infectious virus was defined as E gene reverse transcription (RT)-PCR cycle threshold (Ct) value ≥25. Results: During the 6-month study period, a total of 43 children with 44 episodes of COVID-19 were included. Of the 44 episodes, the median age of the patients included was 8 years old (interquartile range [IQR], 4.9-10.5), and the most common underlying disease was acute lymphoid leukemia (n=30, 68.2%), followed by patients post-hematopoietic stem cell transplantation (n=8, 18.2%). Majority of the patients had mild COVID-19 (n=32, 72.7%), and three patients (7.0%) had severe/critical COVID-19. Furthermore, 2.3% (n=1) died of COVID-19 associated acute respiratory distress syndrome. The largest percentage of the patients showed E gene RT-PCR Ct value ≥25 between 15-21 days (n=13, 39.4%), followed by 22-28 days (n=10, 30.3%). In 15.2% (n=5), E gene RT-PCR Ct value remained <25 beyond 28 days after initial positive PCR. Refractory malignancy status (β, 67.0; 95% confidence interval, 7.0-17.0; P=0.030) was significantly associated with prolonged duration of E gene RT-PCR <25. A patient with prolonged duration of E gene RT-PCR Ct value <25 was suspected to have infectivity shown by the transmission of the virus to his mother at day 86 after his initial positive test. Conclusions: Children that acquire symptomatic COVID-19 during refractory malignancy state are at a high risk for prolonged shedding warranting PCR-based transmission precautions in this cohort of patients.

Antiretroviral Therapy 2006: Pharmacology, Applications, and Special Situations

  • Samuel, Rafik;Bettiker, Robert;Suh, Byung-Se
    • Archives of Pharmacal Research
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    • v.29 no.6
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    • pp.431-458
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    • 2006
  • As we approach the completion of the first 25 years of the human immunodeficiency virus(HIV) epidemic, there have been dramatic improvements in the care of patients with HIV infection. These have prolonged life and decreased morbidity. There are twenty currently available antiretrovirals approved in the United States for the treatment of this infection. The medications, including their pharmacokinetic properties, side effects, and dosing are reviewed. In addition, the current approach to the use of these medicines is discussed. We have included a section addressing common comorbid conditions including hepatitis B and C along with tuberculosis.