• The Journal of Korean Society of Virology
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• v.27 no.1
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• pp.49-57
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• 1997

We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.

• Journal of Veterinary Science
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• v.24 no.3
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• pp.40.1-40.6
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• 2023

Analysis of the VP1 gene sequence of the foot and mouth disease virus (FMDV) is critical to understanding viral evolution and disease epidemiology. A standard set of primers have been used for the detection and sequence analysis of the VP1 gene of FMDV directly from suspected clinical samples with limited success. The study validated VP1-specific degenerate primer-based reverse transcription polymerase chain reaction (RT-PCR) for the qualitative detection and sequencing of serotype O FMDV lineages circulating in India. The novel degenerate primer-based RT-PCR amplifying the VP1 gene can circumvent the genetic heterogeneity observed in viruses after cell culture adaptation and facilitate precise viral gene sequence analysis from clinical samples.

• Pediatric Infection and Vaccine
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• v.16 no.1
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• pp.47-53
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• 2009

Purpose : This study was performed to investigate the serotype distribution of group B streptococcus (GBS) isolated from pregnant Korean women using molecular methods. Methods : The study materials included 42 GBS isolates obtained from the vagina and anorectum of pregnant women in Goyang, Korea between 2005 and 2006. Four clinical isolates with known serotypes (Ia, Ib, III, and V) were used for validation of molecular serotyping. We used serotype-specific primers for identification of the serotypes (Ia, Ib, III, V, and VI). To determine the ambiguous serotypes by serotype-specific PCR, sequence analysis of the PCR amplicons which had been amplified with GBS-common primers was used. Results : The serotypes determined by the molecular methods agreed with the previously known 4 serotypes (Ia, Ib, III, and V). The serotypes of all 42 isolates were successfully determined by molecular methods. The distribution of the GBS serotype was as follows in order of frequency: serotype III was found in 12 isolates (28.6%), serotype V was found in 11 isolates (26.2%), serotype Ia was found in 11 isolates (26.2%), serotype VI was found in 4 isolates (9.5%), serotype Ib was found in 2 isolates (4.8%), and serotype II was found in 2 isolates (4.8%). Conclusion : Serotypes III, V, and Ia were the most frequently identified serotypes in pregnant Korean women. Molecular serotyping is useful for surveillance of the serotype distribution of GBS in colonized pregnant women and GBS diseases of neonates.

• Microbiology and Biotechnology Letters
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• v.27 no.5
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• pp.359-363
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• 1999

To prevent appearance of resistant mosquitoes against $\delta$-endotoxin of bacillus thuringiensis subsp. israelensis (Bti) in field, a mosquitocidal Bacillus thuringiensis strain 21-2(Bt21-2) producing a new type of $\delta$-endotoxin was isolated. The strain Bt 21-2 belongs to H serotype 43, B. thuringiensis subsp. guiyangiensis (Btg). The $\delta$-endotoxins from the strain Bt 21-2 and the strain Bti were a cuboid shape morphologically, but the $\delta$-endotoxin of the strain Bt 21-2 was composed of 150, 90 and 70kDa proteins on SDS-PAGE, and the antigenicity of $\delta$-endotoxin of the strain Bt 21-2 was different from that of the strain Bti on immunoblot. The $\delta$-endotoxin gene of the strain Bt 21-2 was not amplified with specific primers of $\delta$-endotoxin gene (cry4A and cry4B) of the strain Bti on PCR.

• The Korean Journal of Food And Nutrition
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• v.13 no.1
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• pp.1-5
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• 2000

Detection for heat-labile enterotoxin(LT) of enterotoxigenic Escherichia coli(ETEC, EC81, O148:H28) and enterotoxin of enterotoxigentic Clostridium perfringents type A(CP, NCTC8238, Hobbs serotype 2) by use of a polymerase chain reaction (PCR) assay were positive reaction, which using LT gene-specific primers of ETEC with a detection limit equivalent from 100ng/${\mu}\ell$ to 1 pg of a DNA fragment of 417-bp in EC81 and enterotoxin gene-specific primers of CP with a detection limit equivalent from 100ng/${\mu}\ell$ to 10pg of a DNA fragment of 364-bp in NCTC8238. Detection for a LT gene of ETEC highly appeared 10-fold sensitivity than an enterotoxin gene of CP.

• Korean Journal of Food Science and Technology
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• v.32 no.6
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• pp.1336-1340
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• 2000

Overall prevalence of Y. enterocolitica in frozen foods was 5.6% (35 cases of 624 samples). Seasonal variation of contamination was observed. Isolation rate of Y. enterocolitica from samples collected in the second half of the year was six times higher than those of the first half of the year. Serotype of the isolated Y. enterocolitica was mainly serotype O:5 (9 cases). However, 25 cases of 35 isolates could not be serotyped with antiserum used in this study. The biotype test showed that all isolates were non-pathogenic type 1A. The polymerase chain reaction test with ail gene specific primers also confirmed that pathogenic strains were not found in frozen food isolates.

• Clinical and Experimental Pediatrics
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• v.55 no.11
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• pp.424-429
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• 2012

Purpose: Methods for quick and reliable detection of Streptococcus pneumoniae are needed for the diagnosis of pneumococcal disease and vaccine studies. This study aimed to show that sequential multiplex polymerase chain reaction (PCR) is more efficient than conventional culture in achieving S. pneumoniae -positive results. Methods: Nasopharyngeal (NP) secretions were obtained from 842 pediatric patients admitted with lower respiratory infections at Severance Children's Hospital in Korea between March 2009 and June 2010. For identification and serotype determination of pneumococci from the NP secretions, the secretions were evaluated via multiplex PCR technique with 35 serotype-specific primers arranged in 8 multiplex PCR sets and conventional bacteriological culture technique. Results: Among the results for 793 samples that underwent both bacterial culture and PCR analysis for pneumococcal detection, 153 (19.3%) results obtained by PCR and 81 (10.2%) results obtained by conventional culture technique were positive for S. pneumoniae. The predominant serotypes observed, in order of decreasing frequency, were 19A (23%), 6A/B (16%), 19F (11%), 15B/C (5%), 15A (5%), and 11A (4%); further, 26% of the isolates were non-typeable. Conclusion: As opposed to conventional bacteriological tests, PCR analysis can accurately and rapidly identify pneumococcal serotypes.

• The Journal of Korean Society of Virology
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• v.28 no.2
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• pp.147-155
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• 1998

Hantavirus is a genus of the Bunyaviridae family consisting following serotype groups: Hantaan, Seoul, Puumala, Prospect Hill, Thailand, Belgrade, Thotta palayam, Sin Nombre. Most of Hantavirus group have been associated with many clinically similar disease known collectively as hemorrhagic fever with renal syndrome (HFRS). Hantaan virus is the prototype of the genus hantavirus, originally isolated from Apodemus agrarius. Bat was found as a natural host for Hantaan virus in Lee's lab for the first time. Then, Hantaan-like virus was isolated Hantaan-like virus from bat. To identify hantaviruses that are present in Korea among bats, bats were collected from Jeong-Sun, Won-Joo, Chung-Ju and Hwa-Cheon area, RNA was isolated from lung and serum. RT-PCR was performed with a universal primer from M segment. Nested RT-PCR was carried out to differentiate Hantaan, Seoul and Puumala virus using serotype specific primers. As we expected, Hantaan viruses were detected in bats and Seoul virus was not detected. Interestingly, Puumala viruses were also detected in bats from Won-Ju, but not in other areas. Puumala virus is originally isolated from Clethrinomys glareolus, and cause light HFRS. Recently, Paradoxomis webbiana, a wild bird turn out to be a reservoir for Puumala virus in Korea. These data indicate that bat is a new natural reservoir of Puumala virus.

• Korean Journal of Microbiology
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• v.48 no.3
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• pp.175-179
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• 2012

Adenoviruses are an important cause of respiratory tract infections, particularly in infants, young children, and immuno-compromised patients. In this study, we investigated the characteristics of adenoviruses isolated from outpatients with acute respiratory illness in Gyeonggi province of South Korea during 2009-2011. Adenoviruses were detected in 102 of 1,622 (6.3%) specimens by using PCR or real-time PCR with viral specific primers. 76 isolates were obtained from 102 specimens using the A549 cells. Serotypic distributions of isolated adenovirus were analyzed by sequencing of hexon gene. Six different serotypes were identified, which included adenovirus serotypes 1-6. Adenovirus 3 (n=40, 52.6%) was the predominant serotype. The predominant types of adenovirus every year were serotypes 1 and 3 in 2009, serotype 3 in 2010, and serotype 5 in 2011, respectively. Adenoviruses 1, 2, 4, 5, and 6 were isolated sporadically throughout the study period. Adenovirus 3 was present both during outbreaks and in sporadic cases. These results indicate that adenovirus 3 played major causative agent of adenovirus outbreaks in Gyeonggi province of South Korea during 2009-2011. Continuous surveillance for specific serotypes of adenovirus that can cause outbreaks is important.

• Korean Journal of Veterinary Research
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• v.44 no.3
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• pp.399-405
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• 2004

Haemophilus somnus, Mycoplasma bovis and Pasteurella multocida were responsible for respiratory diseases in bovine. Methods for identifying these bacteria had poor sensitivity and specificity. In this paper, PCR assays were applied for rapid identification of H. somnus, M. bovis, P. multocida B:2 and P. multocida capsular types. The specific PCR products were amplified from H. somnus, but not from other bacteria. Ten-fold diluted H. somnus were mixed with P. multocida and then the mixed cultures were inoculated on agar plates. After incubation, PCR was performed with harvest from agar plates and could detect as few as 3.4 CFU/ml of H. somnus. The primers MboF and MboR produced an amplification product unique to M. bovis and sensitivity of PCR was as low as 100 pg of DNA. Only serotype B:2 of P. multocida, the causal agent of haemorrhagic septicemia in bovine, was specifically amplified in PCR among the 16 reference serotypes. The multiplex capsular PCR typing for P. multocida was produced the P. multocida specific product as well as the capsular serogroup-specific product. The present PCR assays should be useful for the rapid identification of bacterial pathogens from bovine respiratory diseases.