Follicular dendritic cell (FDC) sarcoma is rare and is classified either as conventional type or inflammatory pseudotumor (IPT)-like variant. Extranodal presentation is uncommon and nearly all gastrointestinal FDC tumors are of the conventional type. IPT-like variant tumors occur almost exclusively in the liver and spleen and are consistently associated with Epstein-Barr virus (EBV). Here we report the case of a 78-year-old woman with an IPT-like FDC sarcoma presenting as a pedunculated colonic polyp. Histologically, scanty atypical ovoid to spindle cells were mixed with a background of florid lymphoplasmacytic infiltrate, which led to an initial misdiagnosis of pseudolymphoma. These atypical cells expressed CD21, CD23, CD35, and D2-40, and were positive for EBV by in situ hybridization, confirming the diagnosis. The patient was free of disease five months after polypectomy without adjuvant therapy. Although extremely rare, the differential diagnosis for colonic polyp should include FDC sarcoma to avoid an erroneous diagnosis. A review of the 24 cases of IPT-like FDC sarcoma reported in the literature reveal that this tumor occurs predominantly in females with a predilection for liver and spleen, and has a strong association with EBV.
Aged ; Colonic Polyps* ; Dendritic Cell Sarcoma, Follicular* ; Dendritic Cells, Follicular ; Diagnosis ; Diagnosis, Differential ; Diagnostic Errors ; Female ; Granuloma, Plasma Cell ; Herpesvirus 4, Human ; Humans ; In Situ Hybridization ; Liver ; Pseudolymphoma ; Sarcoma ; Spleen ; Taiwan

BACKGROUND: Biomedical products such as viral vaccines can be contaminated with hazardous viruses during manufacturing processes and storage, thus causing harmful side effects. To assure the safety of biomedical products, highly effective and sensitive methods should be available to detect contaminating viruses. In this study, we performed recovery tests to determine the limit of detection of HIV-1. METHODS: An HIV-1 plasmid preparation was serially diluted and spiked into various culture media (DMEM, RPMI-1640, IMDM, GICM, and SDM) containing 10% fetal bovine serum (FBS). The HIV-1 plasmid was detected by PCR alone or a combination of PCR and ELISA (PCR/ELISA). RESULTS: When spiked into DMEM, RPMI, and IMDM, less than 4x10(-2) ng of HIV-1 plasmid was not detectable as HIV-1 PCR products in agarose gel. Intra- and inter-assays (n=6) showed that the PCR-ELISA system could detect PCR products diluted as much as 1, 875 times from HIV-1 plasmid serially spiked in various media. CONCLUSIONS: The PCR/ELISA system can be useful for the detection of trace amounts of hazardous viruses which may be present as contaminants in biological products.
Biological Products ; Culture Media ; Enzyme-Linked Immunosorbent Assay ; HIV-1* ; Limit of Detection ; Plasmids ; Polymerase Chain Reaction ; Sepharose ; Viral Vaccines

No Abstract Available.
Korea* ; Mumps virus* ; Mumps*

Commercial interests towards insect cell cultures have greatly been increased recently, in part due to the widespread use of insect virus-based vectors for efficient expressions of foreign proteins. Insect cell-derived biotechnology products should be free of adventitious agents such as arboviruses and mycoplasmas. The objective of this study was to establish techniques for the viral safety evaluation of insect cell-derived biotechnology products using Japanese encephalitis virus (JEV) as a model, since JEV is a member of arthropod-borne flaviviruses which has been known to be infectious to insect cells such as Sf9 cells. Quantitative assays for viral infectivity, concentrations of an antigen or a genome are a prerequisite for the studies of viral clearance validation. Here, we report the development of a quantitative assay for JEV RNA using real-time reverse transcription-polymerase chain reaction (RT-PCR). The assay was performed using LightCycler and RNA amplification kit SYBR Green I. Viral RNA extracted from stock suspension of JEV with known infectivity titer was made to 10-fold serial dilution, and each dilution was subjected to the assay for the generation of a standard curve. A JEV specific primer was selected from 3' untranslated region with the expected band size of 323 base pairs. The real-time RT-PCR assay resulted in a successful amplification within 5 log dilution ranges of JEV RNA samples, and the sensitivity of the assay was calculated to be approximately 15 TCID50 per reaction. Highly reproducible standard curves were obtained from experiments performed on three different days. The real-time RT-PCR assay for the quantification of JEV will be an efficient alternative tool for viral clearance validation studies of insect cell-derived biotechnology products.
3' Untranslated Regions ; Arboviruses ; Asian Continental Ancestry Group* ; Base Pairing ; Biotechnology* ; Cell Culture Techniques ; Encephalitis Virus, Japanese* ; Encephalitis, Japanese* ; Flavivirus ; Genome ; Humans ; Insects* ; Mycoplasma ; RNA ; RNA, Viral ; Sf9 Cells

As viral vaccines sometimes induce the side effects which intimidate humans, it is urgently required to study about the side effects of viral vaccines. Most vaccines are derived from the biological forms. Hence it should be established the efficient and sensitive evaluation methods for the possibility of dangerous contaminants such as viruses and the stability of vaccines. A lots of vaccine products have been imported, or in sale or being developed in nation. Some of products may be mixed with the hazardous viruses derived from animal or human resources. Such hazardous viruses should be identified specifically and efficiently. Biological products are not permitted to distribute or obtain without presenting the result of absence experiment of hazardous viruses and prion and the validation data of inactivation experiment during processes. In order to detect viruses, there were TEM for viral particles, infectivity assays and detection methods for nucleic acids. However, these methods have defects such as insensitivity and inaccuracy. It should be considered to detect only the hazardous viruses with specificity, precision and accuracy during the processes of preparation, transportation and storage. It is noted to increase the detection limit in order to detect the minute hazardous factors during preparation processes for viral vaccines. This study is focused on the establishment of various sensitive PCR/ELISA methods for HBV virus which enhance the detection limit in order to detect the minute hazardous factors during preparation processes for viral vaccines.
Animals ; Biological Products ; Commerce ; Humans ; Limit of Detection ; Mass Screening* ; Nucleic Acids ; Sensitivity and Specificity ; Transportation ; Vaccines ; Viral Vaccines ; Virion

One-step real-time reverse transcription-polymerase chain reaction (RT-PCR) assay using the MagNA Pure LC and LightCycler(TM) system was developed and validated for the detection and quantitation of hepatitis A virus (HAV) RNA. The assay was evaluated using in-house synthetic HAV RNA standard. The real-time RT-PCR assay could quantitate a dynamic range of HAV RNA standard between 10(2) and 10(8) copies per reaction. The regression coefficient of the standard curve was an 0.99. The detection limit of the assay was 31.3 RNA copies per reaction. The coefficient variations (CVs) of the assay in combination with automated RNA extraction were less than 1.91% in both intra- and inter-assay. The real-time RT-PCR assay for quantitative detection of HAV would serve a useful method for improving the safety of biological products.
Biological Products ; Hepatitis A virus* ; Hepatitis A* ; Hepatitis* ; Limit of Detection ; RNA*

Risk of viral contamination is one of major concerns common to all biologics derived from cultivated cells. Bovine viral diarrhoea virus (BVDV) has widely been known as a contaminant of cell culture-derived vaccines. The objective of the study was to assess the limit of detection and range of quantitation of the detection methods for BVDV using a reverse transcription-polymerase chain reaction (RT-PCR) assay, real-time RT-PCR assay, and RT-PCR-ELISA. One milliliter of cell culture supernatant containing 106.5+/-0.2 median tissue culture infectious dose (TCID50)/ml of BVDV NADL strain was subjected to RNA isolation. The isolated RNA was 10-fold serially diluted and each diluted sample (10-1 to 10-6) was subjected to RT-PCR on a GeneAmpR PCR System 9700 and/or LightCycler(TM). The amplified products were analyzedly (1) agarose gel electrophoresis for RT-PCR assay, (2) melting curve analysis for real-time RT-PCR assay (in this case a program is automatically linked to amplification step), and (3) ELISA using capture and detection probes for RT-PCR-ELISA. The limit of detection of the 3 assay methods was equally estimated to be 316 TCID50/ml of starting virus culture supernatant subjected to the assay. The quantitation range of real-time RT-PCR assay and RT-PCR-ELISA was estimated to be from 3.16x105 to 3.16x102 TCID50/ml of starting virus culture supernatant. The overall results suggested that the 3 assay methods for BVDV detection can be reliably applied to evaluate BVDV contamination in biologics derived from cell cultures.
Biological Products* ; Cell Culture Techniques* ; Electrophoresis, Agar Gel ; Enzyme-Linked Immunosorbent Assay ; Freezing ; Limit of Detection* ; Polymerase Chain Reaction ; Reverse Transcription ; RNA ; Vaccines