• 한국영양학회:학술대회논문집
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• 한국영양학회 2003년도 연합학술대회
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• pp.132.2-132.2
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• 2003

• 대한수의학회지
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• 제63권4호
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• pp.41.1-41.6
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• 2023

Many viruses can infect different types of birds, with poultry being the most susceptible. These viral diseases have a direct negative impact on the poultry industry, with significant economic losses. This study examined a group of the most important viruses that infect backyard chickens in 2 specific areas of Basrah Governorate, south of Iraq. The study analyzed avian influenza viruses (AIVs), Newcastle disease virus (NDV), and infectious bronchitis virus (IBV). Two hundred and ninety oropharyngeal swabs, 150 from Abu Al-Khasib and 140 from Shatt Al-Arab regions in the Basrah governorate, were obtained from backyard chickens with clear respiratory signs. The samples were subjected to viral RNA extraction, and the viral nucleic acids were detected using a reverse transcriptase polymerase chain reaction technique. The overall rate of viral infections was 74.8%, which varied depending on the type of virus: 15.8%, 31.3%, and 27.5% for AIV, NDV, and IBV, respectively. The NDV and IBV had much higher infection rates than that of AIV. In addition, the prevalence of AIV in the Shatt Al Arab district was significantly higher than in the Abul Khasib district. Moreover, there were no significant differences between the NDV and the IBV distributions in either of the targeted regions in this study.

• Korean Journal of Radiology
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• 제21권4호
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• pp.505-508
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• 2020

The epidemic of 2019 novel coronavirus, later named as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is still gradually spreading worldwide. The nucleic acid test or genetic sequencing serves as the gold standard method for confirmation of infection, yet several recent studies have reported false-negative results of real-time reverse-transcriptase polymerase chain reaction (rRT-PCR). Here, we report two representative false-negative cases and discuss the supplementary role of clinical data with rRT-PCR, including laboratory examination results and computed tomography features. Coinfection with SARS-COV-2 and other viruses has been discussed as well.

• BMB Reports
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• 제29권3호
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• pp.241-247
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• 1996

The Philadelphia (Ph) chromosome is the single most intensively studied chromosome alteration characterizing a human malignancy. The specific genetic alteration of chronic myelogenous leukemia (CML) is the formation of the BCR/abl fusion gene in leukemic cells. The presence of the BCR/abl gene has important diagnostic and prognostic implications in CML. The detection of BCR/abl transcripts by reverse transcriptase based polymerase chain reaction (RT-PCR) was investigated in patients with CML in whom the Ph chromosome abnormality was documented by cytogenetic analysis. In a total of 68 CML patient cases, the Ph chromosome was found in 53 cases (77.9%) by cytogenetic analysis. On the other hand, sixty two cases (91.2%) were detected to have BCR/abl gene rearrangement Of these, b3a2 was 44 cases (64.7%) and b2a2 was 17 cases (25,0%). There was one case with both b3a2 and b2a2 (1.5%). Of the fifteen cases of Ph chromosome negative by cytogenetic anlaysis, the BCR/abl gene was observed in nine cases, The results of BCR/abl fusion gene confirmed by the direct sequencing method correlated well with PCR analysis, The amplified PCR products were detected by $1{\times}10^{-5}$ dilutions. In conclusion, PCR technique is sensitive, rapid and relatively simple for a laboratory test in detecting the BCR/abl fusion gene with CML regardless of the result of cytogenetic analysis.

• Journal of Plant Biotechnology
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• 제5권2호
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• pp.83-86
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• 2003

A reverse transcription polymerase chain reaction (RT-PCR) protocol was developed using two specific 22-mer primers located in coat protein gene of SPFMV. A 411 bp PCR-product was detected in virus infected plants as well as tissue culture raised sweet potato but not in healthy plants. For optimization of RT-PCR protocol, the optimum crude nucleic acid concentration, annealing temperature, primer concentration and numbers of PCR-cycle for maximum sensitivity and specificity were determined. The optimum condition for RT-PCR was as follows: RT-PCR reaction mixture was one-step mixture, containing 50 pmol of primer, 30 units of reverse transcriptase, 5 units of RNasin, and the crude nucleic acid extracts (200 ng). In RT-PCR, cDNA was synthesized at $42^{\circ}C$ for 45 min before a quick incubation on ice after pre-denaturation at $95^{\circ}C$ for 5 min. The PCR reaction was carried out for 40 cycles at $96^{\circ}C$ for 30 see, $63^{\circ}C$ for 30 sec, $72^{\circ}C$ for 1 min, and finally at $72^{\circ}C$ for 10 min. The viral origin of the amplified product was confirmed by sequencing, with the sequence obtained having $95-98\%$ homology with published sequence data for SPFMV. The benefits of this RT-PCR based detection of SPFMV would be simple, rapid and specific.

• Journal of Microbiology and Biotechnology
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• 제4권1호
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• pp.20-23
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• 1994

The polymerase chain reaction was used to study the genes inducible under stress from the heavy metal cadmium. Schizosaccharomyces pombe, grown in the presence or absence of sublethal concentration of cadmium, was isolated to purify the total RNAs. The Induced RNA Random Fishing (IRRF) method in which random oligonucleotides were used as primers was applied to the identification of cadmium-induced gene expressions. A PCR-DNA product of 400-bp was cloned and sequenced. Computer analysis showed that this DNA has no homology with any known DNA sequences in GenBank or EMBL databases. The induction of this gene was confirmed by Northern blot analysis of total RNAs isolated from both cadmium-treated and untreated yeast cells.

• Clinical and Experimental Pediatrics
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• 제53권2호
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• pp.129-135
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• 2010

Human rhinoviruses (HRVs) is a nonenveloped, single stranded RNA virus belonging to the family Picornavirudae. Transmission by direct contact such as hand-to-hand, hand-to-nose, and hand-to-eye has been readily demonstrated in experimental settings. HRV are the most frequent causes of common cold infection, however, they are also known to replicate in the lower respiratory tract and associated with more severe respiratory illnesses such as asthma. New technique such as reverse transcriptase polymerase chain reaction and molecular typing in HRV has been developed and our understanding of the importance of these respiratory viruses. HRVs consisted of 101 serotypes that are classified into groups A and B according to sequence variations. And there is a newly identified set of HRVs, called Group C, and it is currently under investigation. In recent study using PCR techniques, HRVs accounted for approximate 50-80% of common colds and 85 % of childhood asthma exacerbations and in more than half of adult exacerbations. However, the mechanisms of HRV- induced asthma exacerbations are poorly understood. This review discusses the association between HRVs and childhood asthma.

• Journal of Microbiology and Biotechnology
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• 제15권6호
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• pp.1384-1387
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• 2005

Hantavax(R) is one of the killed Hantavirus vaccines, and is commercially available in South Korea. This vaccine was developed by inactivation of virus isolated from infected suckling mouse brain with formalin. Although Hantavax(R) can induce neutralizing antibodies in vaccinees, the strength of this induction and the duration of the humoral immune response are controversial issues. In this study, we studied the native conformation of the killed vaccine by antigen-capture reverse transcriptase polymerase chain reaction with patient and vaccinee sera containing neutralizing antibodies against Hantavirus. The results showed that Hantavax(R) could bind HTNV patient and vaccinee sera like live virus, suggesting that the integrity of the viral epitope is maintained in Hantavax(R) and induces the protective antibodies, even though the virus was inactivated with formalin.

• 현장농수산연구지
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• 제5권1호
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• pp.57-63
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• 2003

The bovine rotavirus(BRV), bovine coronavirus(BCV) and bovine viral diarrhea virus(BVDV) are main viruses of bovine viral diarrhea disease. These viruses could be rapidly amplified by the reverse transcriptase polymerase chain reaction(RT-PCR). This study was conducted to develop rapid and accurate diagnostic methods of these viral diseases by multiplex RT-PCR. Specific primers were designed based on the sequences reported by Chang KO et. al. (1997) and Schroeder BA, et. al. (1990), RNA were prepared from the cultured viruses, first-stranded DNAs were synthesised by reverse transcriptase. PCR were conducted to amplify specific regions of the viruses by multiplex. Three bands such as 1,062bp for BRV, 458bp for BCV, and 300bp for BVDV were successfully produced by multiplex RT-PCR. In conclusion, this result suggested that these viruses could be diagnosed rapidly and accurately by multiplex RT-PCR.

• 한국축산식품학회지
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• 제31권2호
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• pp.158-165
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• 2011

A sensitive reverse transcriptase-PCR (RT-PCR) method to detect viable Escherichia coli O157:H7 in milk was established. The primer sets were designed based on the nucleotide sequences of the rfbE (per) and wbdN genes in the O157 antigen gene cluster of E. coli O157:H7. RT-PCR using five different primer sets yielded DNA with sizes of 655, 518, 450, and 149-bp, respectively. All five of the E. coli O157:H7 strains were detected by RT-PCR, but 11 other bacterial species were not. The sensitivity of RT-PCR was improved by adding yeast tRNA as a carrier to the crude RNA extract. The RT-PCR amplifying the 149-bp DNA fragment was the most sensitive for detecting E. coli O157:H7 and the most refractory to the bactericidal treatments. Heat treatment at $65^{\circ}C$ for 30 min was the least inhibitory of all bactericidal treatments. Treatment with RNase A strongly inhibited the RT-PCR of heated milk but not unheated milk. This study described RT-PCR methods that are specific and sensitive with a detection limit of 10 E. coli O157:H7 cells, and showed that pre-treating milk samples with RNase A improved the specificity to detect viable bacteria by RT-PCR.