Evaluation of Seeplex(TM) RV Detection Kit for Detecting Rhinovirus, Human Metapneumovirus, and Coronavirus / 대한진단검사의학회지

BACKGROUND: Direct antigen test (DAT) and culture are primary tests to diagnose infections by respiratory viruses, but are mainly available for the traditional viral pathogens such as respiratory syncytial virus (RSV), influenza virus, parainfluenza virus (PIV), and adenovirus in clinical laboratories. The objective of this study was to evaluate a multiplex reverse transcriptase-PCR method using Seeplex(TM) RV Detection kit (Seegene, Korea) for the detection of rhinovirus, coronavirus, and human metapneumovirus (hMPV). METHODS: From January to May 2007, nasopharyngeal aspirates (NPAs) from pediatric patients negative for culture and DAT of traditional viral pathogens were tested with Seeplex(TM). All the amplicons were directly sequenced and homology of the sequences was searched in the National Center for Biotechnology Information (NCBI) database. Patients' medical records were reviewed for clinical and demographic features. RESULTS: Forty-seven (26.4%) of 178 NPAs were positive: 18 rhinovirus, 15 hMPV, 4 RSV A, 3 coronavirus OC43, 3 influenza virus A, 2 adenovirus, 1 coronavirus NL63, and 1 RSV B. Based on maximum identity, each of the sequences indicating rhinovirus, hMPV, and coronavirus OC43 matched to the corresponding viruses with homology of 94-98%, 96-99%, and 98-100%, respectively. Seeplex(TM) positive patients were 0-11 yr old with a male:female ratio of 1.5:1. Clinical diagnoses included 9 pneumonia, 6 bronchiolitis, 2 cold, 1 asthma exacerbation for rhinovirus; 10 pneumonia, 4 bronchiolitis, and 1 clinical sepsis for hPMV; and 1 pneumonia, 2 croup, and 1 cold for coronavirus. CONCLUSIONS: Multiplex reverse transcriptase-PCR method using Seeplex(TM) RV Detection kit is a reliable test to detect rhinovirus, hMPV, and coronavirus. It may improve the diagnostic sensitivity for RSV, influenza virus, PIV, and adenovirus.

Seroepidemiological Survey of Hantavirus Infection of Wild Rodents Trapped from 1994 to 1998 in Korea

Hantaviruses belong to the genus Hantavirus and Hantaan, Seoul, Puumala, Belgrade and Sin Nombre viruses are the etiolgic agents of two serious hantaviral diseases of humans. The rodent hosts and the specific etiologic agents of haemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS) are known and many reported cases occurred in Eurasia and Americas. Wild rodents trapped in 13 different areas of Korea from 1994 to 1998 were investigated against hantavirus infection. A total of 718 wild rodents and 10 species were trapped and found 630 (87.7%) of them were Apodemus agrarius. Indirect immunofluorescent antibody technique (IFAT) was performed for hantaviruses infections using different hantavirus antigens. Hantavirus antibodies were found in 68 (10.8%) out of 630 A. agrarius, 8 (42.1%) of 19 Rattus norvegicus. Among 68 lungs and other tissues of antibody positive A. agrarius, 5 (7.4%) were antigen positive. IFA titers of 5 positive A. agrarius sera showed higher titers against Puumala or Sin Nombre viruses than Hantaan virus. These results suggest that there may be are possibilities of existence of a noble hantavirus in Korean wild rodents.

Murine Typhus as a Cause of Acute Respiratory Distress Syndrome in Endemic Area / 결핵

BACKGROUND: Murine typhus is a fea-borne, worldwide Rickettsial disease caused by Rickettsia typhi. Its symptoms are typically mild byt sometimes can be fatal. The major clinical features include fever, rash, and headache. Recently, we experienced 6 cases of ARDS associated with a Rickettsia typhi infection. This study was aimed to analyze the attributing factors for fatal murine typhus and to review the characteristics of the pateints who showed acute respiratory distress syndrome as the initial presentation. METHODS: The medical records of 15 patients diagnosed as murine typhus were reviewed. The diagnosis was made by single titers of 1:512 or higher, or a 4-fold rise with compatible clinical features. Acute Respiratory Distress syndrome (ARDS) was define according to the American-European Consensus Conference. The characteristics between the ARDS group and the non-ARDS group of murine typhus were compared. RESULTS: Six patients developed ARDS as their initial presentation. Two of them were women and three of them had lived urban area. None of them a showed skin rash. One of them expired during treatment. The time lapse until the commencement of the specific treatment, the lower serum albumin level, the higher serum total bilirubin level, the higher APACHE III score and the higher MOD score were significantly associated with the ARDS group compared to the non-ARDS group. CONCLUSIONS: Murine typhus should be considered as one of the etiologies for the ARDS of unknown cause, particularly in an endemic regions. ARDS caused by Murine typhus generally has a good prognosis.

Rapid Detection and Identification of Human Respiratory Syncytial Virus, Human Parainfluenza Virus Type 1, 2 and 3 by Single-tube Multiplex Reverse Transcription Polymerase Chain Reaction

Laboratory diagnosis of respiratory viral infection has traditionally been based upon virus isolation and/or viral antigen identification. Recently, more sensitive and specific nucleic acid detection methods by reverse transcription- polymerase chain reaction (RT-PCR) have been developed, however, conventional RT-PCR can identify only a single suspected virus. To identify the causative agents which belong to Paramyxoviridae of respiratory virus infections, we have developed a single-tube multiplex RT-PCR using four primer sets which can amplify respiratory syncytial virus and parainfluenza virus type 1, 2 and 3 simultaneously. Assay sensitivity of single-tube multiplex RT-PCR allowed a detection in the range of 3~500 TCID50 and there were no cross amplification among other respiratory viral agents based on the test using reference virus stocks. The single-tube multiplex RT-PCR was able to directly detect viruses in respiratory specimens, with virus being detected 11 of 80 samples as compared to 9 of 80 samples detected by indirect immunofluorescence or antigen detection following shell vial culture. This result suggests that the single-tube multiplex RT-PCR can be established as a more sensitive and rapid diagnostic application than shell vial assay for the detection of respiratory infection of Paramyxoviridae.

Simultaneous Detection and Identification of Human Respiratory Syncytial Virus, Influenza Virus A ( H3N2 , H1N1 ) and B by One - tube Multiplex Reverse Transcription Polymerase Chain Reaction

Respiratory syncytial virus (RSV) and Influenza virus are the most common pathogen for causing severe upper respiratory infection in all age groups. A multiplex reverse transcription polymerase chain reaction (RT-PCR) has been developed to detect and subtype influenza A (H3N2 and H1N1), B virus and RSV simultaneously in one tube reaction. Amplification with primers derived from conserved sequences within the nucleocapsid for RSV and hemagglutinin subunit for Influenza A (H3N2 and H1N1) and B viruses yielded a 384 bp, a 300 bp, a 236 bp and a 151 bp, respectively. Assay specificity was confirmed by pulse field gel electrophoresis and autosequencing method. Assay sensitivity was 3 PFU/ml of RSV, 22 PFU/ml, 45 PFU/ml of Influenza type A (H3N2 and H1N1) and 6.6 PFU/ml of Influenza B virus by plaque assay. A rapid and sensitive detection method of a one-tube with multiplex RT-PCR capable of identifying more than one viral template as well as synchronizing reverse transcription and PCR had the potential to produce considerable savings of time and cost effectiveness in the diagnostic laboratory.

Seroepidemiologic Survey of Haemorrhagic Fever With Renal Syndrome from 1994 till 2000

No abstract available.

Molecular Characterization of Hantavirus Isolated from Bandicota indica Captured in Indonesia and Thailand

No Abstract Available.

An Immunoserological Study of Vaccine Against Haemorrhagic Fever with Renal Syndrome

Since HantavaxTM, formalin inactivated Hantaan virus vaccine (10,240 ELISA units/ml), has been developed in 1990 to prevent against haemorrhagic fever with renal syndrome (HFRS) caused by Hantaan or Seoul virus, it has been commercially available in Korea. Twenty-one healthy people were booster shot once and twice after primary basic vaccination with HantavaxTM. Seroconversion rates were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA), and plaque reduction neutralization test (PRNT). Seroconversion rates of 21 vaccinees at one year after primary basic vaccination were 52.3%, 95.2%, 0.0%, 47.6%, and 28.6%, and 13 vaccinees of one month after 1st booster vaccination were 100%, 100%, 30.7%, 100% and 100% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates declined slightly by twenty months, and they were 84.6%, 92.3%, 0.0%, 84.6% and 69.2% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates of 9 vaccinees at three months after 2nd booster vaccination were 100%, 100%, 0.0%, 100%, and 88.9%, and 16 vaccinees at one year after the 2nd booster vaccination were 87.5%, 93.8%, 0.0%, 87.5% and 81.3% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Based on the above result HantavaxTM has proved a vigorous anamnestic response after the 1st and the 2nd booster vaccination and has persisted higher fluorescence, agglutination and neutralizing antibody titers in vaccinees.

Epidemiological Analysis of Viral Respiratory Infections and Comparison of Isolation Rate of Various Clinical Specimens / 감염

BACKGROUND: We studied the epidemics of respiratory viral infections in Korea and examined various respiratory tract specimens for the presence of respiratory viruses, since the accuracy of rapid detection method depends, in part, on the source of the specimens. METHODS: Over a 24-month period, from March 1997 through February 1999, a total of 1,574 clinical specimens were submitted for the detection of respiratory viruses. A shell vial technique with commercially available monoclonal antibodies directed against respiratory viruses was used to detect respiratory syncytial virus, adenovirus, influenza virus, and parainfluenza virus in clinical specimens, which included throat swab, nasopharyngeal aspirate, tracheal aspirate, and bronchoalveolar lavage (BAL) fluid. RESULTS: Overall positive rate of respiratory viruses was 73/1574 (4.6%). Respiratory viruses were predominantly found between December and February. High incidences were observed among those younger than 2 years and those older than 50 years. The numbers of viral isolates were 3/69 (4.3%) for throat swab, 26/459 (5.7%) for nasopharyngeal aspirate, 11/315 (3.2%) for tracheal aspirate, and 30/528 (5.7%) for BAL fluid. CONCLUSIONS: Nasopharyngeal aspirate and BAL fluid appear to permit increased detection of the respiratory viruses compared with throat swab or tracheal aspirate. However, throat swab may be good specimen for the detection of influenza virus and parainfluenza virus.

A Field Efficacy Trial of Inactivated Hantaan Virus Vaccine (Hantavax(TM)) Against Hemorrhagic Fever with Renal Syndrome (HFRS) in the Endemic Areas of Yugoslavia from 1996 to 1998

In Yugoslavia, homorrhagic fever with renal syndrome (HFRS) is one of the important national health problem, but no vaccine has been used to prevent HFRS. Since first HFRS case in 1952, sporadic cases of HFRS occurred every year and over 4,000 registered cases with 1~16% mortality so far. We performed a prospective, randomized double-blind placebo-controlled trial to evaluate the effectiveness of Hantavax(TM) against HFRS in 3,900 healthy adults living in the endemic areas of Yugoslavia. 1,900 people were given 0.5 ml of Hantavax subcutaneously twice at one month interval and a booster shot at one year after. For controls other 2,000 healthy people were given 0.5 ml of physiolosical saline as a placebo. We investigated HFRS cases in both the vaccinated and nonvaccinated groups by monitoring the program for patient registration in the areas from 1996 to 1998, and the effect of vaccine was analyzed epidemiologically No confirmed case of HFRS was observed among 1,900 Hantavax vaccinees, while 20 confirmed cases were observed among 2,000 nonvaccinated control group. There were no remarkable side effects among the vaccinees either locally or in general after inoculation of the vaccine. The Hantavax vaccine showed statistically significant protective efficacy against HFRS among Yugoslavian people.

A Field Efficacy Trial of Inactivated Hantaan Virus Vaccine (Hantavax(TM)) Against Hemorrhagic Fever with Renal Syndrome (HFRS) in the Endemic Areas of Yugoslavia from 1996 to 1998

In Yugoslavia, homorrhagic fever with renal syndrome (HFRS) is one of the important national health problem, but no vaccine has been used to prevent HFRS. Since first HFRS case in 1952, sporadic cases of HFRS occurred every year and over 4,000 registered cases with 1~16% mortality so far. We performed a prospective, randomized double-blind placebo-controlled trial to evaluate the effectiveness of Hantavax(TM) against HFRS in 3,900 healthy adults living in the endemic areas of Yugoslavia. 1,900 people were given 0.5 ml of Hantavax subcutaneously twice at one month interval and a booster shot at one year after. For controls other 2,000 healthy people were given 0.5 ml of physiolosical saline as a placebo. We investigated HFRS cases in both the vaccinated and nonvaccinated groups by monitoring the program for patient registration in the areas from 1996 to 1998, and the effect of vaccine was analyzed epidemiologically No confirmed case of HFRS was observed among 1,900 Hantavax vaccinees, while 20 confirmed cases were observed among 2,000 nonvaccinated control group. There were no remarkable side effects among the vaccinees either locally or in general after inoculation of the vaccine. The Hantavax vaccine showed statistically significant protective efficacy against HFRS among Yugoslavian people.

Immune Response and Antibody Persistence against Hantaan Virus of Vaccinees with Hantavax(TM) / 감염

BACKGROUND: Hantavax(TM) was developed for preven-tion of hemorrhagic fever with renal syndrome caused by Hantaan or Seoul virus in 1990, and has been commer-cially available in Korea since then. Because Hantavax (TM) has such short usage history, the duration of antibody persistency in vaccinees has not been well studied. METHODS: 61 healthy people were immunized subcu-taneously with Hantavax (TM) twice at one month intervals as primary vaccination. 21 vaccinees were tested at 1 ~4 months after primary vaccination and 40 vaccinees were tested at one year after primary vaccination and then one month and 1 ~2 years after booster vaccination. Antibody titers were measured by immunofluorescent assay(IFA), Hantaan virus antigen-coated high density particle agglu-tination assay(HDPA), and plaque reduction neutralization test(PRNT). RESULTS: Seroconversion rates of 21 vaccinees at 1 ~ 4 months after primay vaccination were 20/21(95.2%), 19/21(90.5%) and 14/21(66.7%); seropositivity of 40 vaccinees at one year after primary vaccination was 25/40 (62.5%), 18/40(45.0%), and 9/40(22.5%) by IFA, HDPA, and PRNT, respectively. Seroconversion rates of 8 vaccinees at one month after booster vaccination were 8/ 8(100 %), 8/ 8(100%); antibody persistence rate of 11 vaccinees at 20 months after booster vaccination were 11/ 12 (91.7%), 9/ 12(75.0%), and seroconversion rates of 7 vaccinees at 3 months after second booster vaccination were 7/7(100%) and 6/7(85.7%) by IFA and PRNT, respectively. Geometric mean antibody titers of 21 vaccinees at 1-4 months after primary basic vaccination were 262, 248, 120; and those of 40 vaccinees at one year after primary vaccination were 90, 56, and 24 by IFA, HDPA, and PRNT, respectively. Geometric mean antibody titers of 8 vaccinees at one month after booster vaccination were 852, 183, of 12 vaccinees at 20 months after booster vaccination were 296, 33, and of 7 vaccinees at 3 months after second booster vaccination were 549 and 46 by IFA and PRNT, respectively. CONCLUSION: The booster vaccination is necessary at 12 months after primary vaccination to maintain high levels of antibodies which persist at least two years after booster vaccination.

Isolation of the Hantaviruses from the Lungs of Bandicota indica Captured in Indonesia and Thailand

Various hantaviruses were isolated from HFRS patients and various rodent species, in many parts of the world. Bandicotas were captured at Yogyakarta, east region of Sumatura island, Indonesia; and 4 rodents species including Bandicotas were captured at Chiang Rai in Thailand during 1995. Sera were collected from captured andicotas and other rodent spicies were screened for antibody test against Hantaan (HTN), Seoul (SEO), Puumala (PUU) and Sin Hombre (SN) viruses by immunofluoresence antibody assay (IFA). Hantavirus antigen in lung tissues were tested by IFA. Among 55 captured Bandicota indica in Indonesia, 14 (25.5%) were antibody positive against HTN, SEO, PUU and SN virus. Hantavirus antigen were detected from 5 (9.0%) out of 55 lungs tested. Among 34 captured Bandicota indica in Thailand, 9 (26.5%) were antibody positive against HTN, SEO, PUU and SN virus. Among 34 lungs tissues of Bandicota indica examined, 3 (8.8%) were antigen positive. In other rodent species, antibody positive against Hantaviruses of Rattus rattus, Rattus losea and Mus cervicolor were 4/62(6.5%), 5/25(20%), 1/1(100%), respectively. But no one has antigen in their lung tissues. Antigen positive lungs suspension were inoculated into vero E6 cells for virus isolation and 4 viruses were isolated from Indonesian Badicota and 3 viruses from Thailand.

Detection of Antibodies against Shope Fibroma Virus and Formation of Fibroma in the Korean Domestic Rabbits

In our preliminary study to find antiviral or antitumor agents from Korean natural products, we found that the Shope fibroma virus (SFV) induced fibromas reaching maximum size at 5~6 days with spontaneous disappearance at 15~20 days after SFV intracutaneous inoculation into Korean domestic rabbits. However, the sizes of fibromas of rabbits at day 5 after virus inoculation were significantly different individually. Assuming that the variation of tumor size was due to either susceptibility or the preexisting antibodies against SFV in the Korean domestic rabbits, the rabbits were checked for the antibodies against SFV by IFAT using SFV infected RKl3 cells. The antibody positive rate of normal Korean domestic rabbits was 32.8% and the sizes of the fibromas of the positive rabbits were significantly smaller than those of negative rabbits (p<0.0001). The fibroma sizes were dependent on the antibody titers of rabbits to SFV. The sizes of fibromas after inoculation of SFV into immunized rabbits were about one tenth of those by the first inoculation into normal rabbits. This is the first report on the antibody prevalence against SFV among normal Korean domestic rabbits and it suggest the existence of a wild fibroma virus or related virus in Korea.

Immune Reaction of the Vaccinated Hamsters with Combined Hantaan-Puumala Vaccine

A large number of viruses belonging to Genus Hantavirus in Family Bunyaviridae are etiologic agents for hemorrhagic fever with renal syndrome (HFRS), or hantavirus pulmonary syndrome (HPS). Hantaan (HTN), Seoul (SEO), Belgrade (BEL), Puumala (PUU) serotype viruses are well known causative agents for HFRS in Eurasian continent. Among those viruses Hantaan and Seoul serotypes are well known to cause HFRS in Korea, but there are some sporadic incidence by other than Hantaan or Seoul viruses. Recently we have developed the combined Hantaan-Puumala virus vaccine to prevent world-wide occurring HFRS. This combined vaccine is formalin inactivated, suckling mouse and suckling hamster brain extracts for Hantaan and Puumala viruses, respectively. Protein contents of this purified candidate vaccine is 27 microgram/ml, which contains 1,024 ELISA antigen units to each virus, but content of myelin basic protein which is causing experimental allergic encephalomyelitis is legs than 0.1 ng/ml. Thirty hamsters were given twice at one month interval intra-muscularly and bled on 30 days after each vaccination from retro-orbital sinus vein. Antibody titers were tested against 5 major serotype viruses, Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses by IFA and PRNT. The mean IF antibody titers on 30 days after primary shot were 78.4, 68.8, 68.8, 37.9, and 15.6; mean neutralizing antibody titers were 65.4, 12, 6.1, 65.6 and 0.5 against Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses, respectively. The mean IF antibody titers on 30 days after booster shot were 686.9, 567.5, 550.4, 516.3, and 430.9; and neutralizing antibody titers were 710.8, 41.9, 24.3, 409.9, and 1.6 against Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses, respectively.

The persistence of immunity against hemorrhagic fever with renal syndrome among Hantaan virus vaccinees

No abstract available.

The persistence of immunity against hemorrhagic fever with renal syndrome among Hantaan virus vaccinees

No abstract available.

Persistence of antibodies after immunization with the inactivated vaccine against hemorrhagic fever with renal syndrome in humans

No abstract available.

A study of immunogenicity of an inactivated vaccine againsthemorrhagic fever with renal syndrome

No abstract available.