ABSTRACT
A 70-year-old man presented with lower back pain and cyanotic changes in his left lower extremity. He was diagnosed with infected aortic aneurysm and infectious spondylitis. He had received intravesical Bacillus Calmette-Guerin (BCG) therapy up to 1 month before the onset of symptoms. The aneurysm was excised and an aorto-biiliac interposition graft was performed. Mycobacterium tuberculosis complex was cultured in the surgical specimens. Real-time polymerase chain reaction (PCR) targeting the senX3-regX3 region, and multiplex PCR using dual-priming oligonucleotide primers targeting the RD1 gene, revealed that the organism isolated was Mycobacterium bovis BCG. The patient took anti-tuberculosis medication for 1 year, and there was no evidence of recurrence at 18 months follow-up.
Subject(s)
Aged , Humans , Administration, Intravesical , Aneurysm , Aneurysm, Infected , Aortic Aneurysm , Bacillus , DNA Primers , Follow-Up Studies , Genes, rev , Low Back Pain , Lower Extremity , Multiplex Polymerase Chain Reaction , Mycobacterium bovis , Mycobacterium tuberculosis , Mycobacterium , Real-Time Polymerase Chain Reaction , Recurrence , Spondylitis , Transplants , Urinary Bladder Neoplasms , Urinary BladderABSTRACT
<p><b>OBJECTIVE</b>To develop a safe and effective lentivirus vaccine model and provide insights into the development of other lentivirus vaccines.</p><p><b>METHODS</b>In this study, a construct of pGPT was made by deleting env gene in the infectious Equine infectious anemia virus (EIAV) molecular clone of WU57. Since the overlaping of EIAV Rev gene with env gene, there was no Rev gene in the construct of pGPT. For compensation of Rev function, the construct of pGPTC was made by inserting 4 copies of constitutive RNA transport elements (CTEs) from Mason-Pfizer monkey virus into the construct of pGPT. In addition, a construct designated pTEB expressing EIAV Env protein was made while env gene-minus viruses were made by co-transfection of pGPT/pTEB or pGPTC/pTEB into 293 cells. Western blot was used to identify the development of recombinant virus particles. Then immunofluorescence assay was used to evaluate the infectivity of recombinant virus particles in vitro.</p><p><b>RESULTS</b>EIAV proteins expression was detected in the supernatant of transfected 293 cells by Western blot within pGPTC/pTEB transfected cells. However, no evidence of EIAV proteins expression was observed within pGPT/pTEB transfected cells. EIAV proteins expression was detected in the first round but not in the second round infected EK cells with EIAV(GPTC) by immunofluorescence assay.</p><p><b>CONCLUSION</b>Rev/RRE was necessary for expression of viral structural proteins; CTEs from Mason-Pfizer monkey virus was functionally interchangeable with EIAV Rev/RRE to help RNAs transportation out of nucleus to express structural proteins and EIAV particles were produced in the transfected 293 cells. A live EIAV recombinant virus with single round infection had been developed.</p>
Subject(s)
Animals , Humans , Blotting, Western , Cells, Cultured , Fluorescent Antibody Technique , Genes, rev , Haplorhini , Horses , Infectious Anemia Virus, Equine , Genetics , Lentivirus , Allergy and Immunology , Lentivirus Infections , Allergy and Immunology , Mason-Pfizer monkey virus , Genetics , Transfection , Vaccines, Synthetic , Viral Vaccines , Genetics , Allergy and ImmunologyABSTRACT
BACKGROUND: Among Enterobacter spp. isolates from clinical specimens in Korea, the incidence of resistance to expanded-spectrum cephalosporins is becoming an ever-increasing problem. This study was designed to determine the prevalence of expanded-spectrum cephalosporins-resistant Enterobacter spp. isolates from patients in a tertiary care hospital in Busan, Korea, and to characterize the mechanism of resistance. MATERIALS AND METHODS: Nonduplicated clinical isolates of Enterobacter spp. were collected during the period of 1999-2000 in Kosin Medical Center, Busan, Korea. Antimicrobial susceptibilities were tested by disk diffusion method. Cefotaxime-resistant or intermediate isolates were examined for extended-spectrum beta-lactamase (ESBL)-production by double disk synergy (DDS) test. Minimal inhibitory concentrations were determined by agar dilution method. For detection of blaTEM and blaSHV genes, polymerase chain reactions (PCRs) were performed, and the DNA sequences of blaTEM and blaSHV genes were determined by using dideoxy-chain termination method. RESULTS: From 1999 to 2000, a total of 306 Enterobacter spp. strains were isolated from patients in Kosin Medical Center. Forty one percents of Enterobacter spp. isolates were susceptible to cefotaxime. Among 90 isolates resistant or intermediate to cefotaxime, 26 isolates (29%) showed positive results in double disk synergy test. Among DDS-positive- isolates, 22 isolates contained both of blaTEM and blaSHV genes, while one isolate only contained blaTEM gene and two isolates only contained blaSHV gene. Among 64 DDS-negative isolates, 47 isolates contained blaTEM genes, and 12 isolates also contained blaSHV genes. Nucleotide sequence analysis of PCR products from 10 DDS-positive and 6 DDS-negative isolates, which contained both of blaTEM and blaSHV genes, revealed that blaTEM-1b and blaSHV-12 genes were the dominant types of beta-lactamase gene. CONCLUSION: Expanded-spectrum cephalosporins-resistant Enterobacter spp. were wide spread in Kosin Medical Center, Busan, Korea. Some of the resistant isolates acquired resistance by production of ESBLs, and blaSHV-12 gene was the most frequent ESBL gene in cefotaxime-resistant Enterobacter spp.
Subject(s)
Humans , Agar , Base Sequence , beta-Lactamases , Cefotaxime , Cephalosporins , Diffusion , Enterobacter , Genes, rev , Incidence , Korea , Polymerase Chain Reaction , Prevalence , Tertiary HealthcareABSTRACT
Phylogenetic analysis was conducted to monitor transmission of HIV and to investigate the genetic structure of primary isolates from 12 HIV-1 subtype A infected Koreans. The individuals infected with subtype A viruses had been diagnosed as HIV-1 seropositives during the period 1987 to 1995 and blood samples have been collected from 1991 to 1997. DNA of each individual was isolated from uncultured or cultured peripheral blood mononuclear cells. V3-V5 (0.7 kb) fragment of HIV-1 rev gene was amplified by nested polymerase chain reaction and the PCR products were sequenced. The mean value of the divergence of nucleotide of HIV-1 euv V3-V5 fragment was 17.0+/-4.06% (8.6~25.8%) within HIV-1 subtype A isolates from Koreans. This diversity was higher than those of African isolates (13.7+/-2.66%). In the phylogenetic tree, Korean subtype A isolates were not grouped together, but intermingled into African isolates. The results of this study suggested that HIV-1 subtype A variants be introduced from multiple sites of Africa into Korea and the big genetic diversity of Korea HIV-1 subtype A isolates may be further influenced by the range of geographic locations in which the infection occurred rather than the elapsed time between infection and collection of samples and the disease progression.
Subject(s)
Africa , Disease Progression , DNA , Genes, env , Genes, rev , Genetic Structures , Genetic Variation , Geographic Locations , HIV , HIV-1 , Korea , Polymerase Chain ReactionABSTRACT
Phylogenetic analysis was conducted to monitor transmission of HIV and to investigate the genetic structure of primary isolates from 12 HIV-1 subtype A infected Koreans. The individuals infected with subtype A viruses had been diagnosed as HIV-1 seropositives during the period 1987 to 1995 and blood samples have been collected from 1991 to 1997. DNA of each individual was isolated from uncultured or cultured peripheral blood mononuclear cells. V3-V5 (0.7 kb) fragment of HIV-1 rev gene was amplified by nested polymerase chain reaction and the PCR products were sequenced. The mean value of the divergence of nucleotide of HIV-1 euv V3-V5 fragment was 17.0+/-4.06% (8.6~25.8%) within HIV-1 subtype A isolates from Koreans. This diversity was higher than those of African isolates (13.7+/-2.66%). In the phylogenetic tree, Korean subtype A isolates were not grouped together, but intermingled into African isolates. The results of this study suggested that HIV-1 subtype A variants be introduced from multiple sites of Africa into Korea and the big genetic diversity of Korea HIV-1 subtype A isolates may be further influenced by the range of geographic locations in which the infection occurred rather than the elapsed time between infection and collection of samples and the disease progression.
Subject(s)
Africa , Disease Progression , DNA , Genes, env , Genes, rev , Genetic Structures , Genetic Variation , Geographic Locations , HIV , HIV-1 , Korea , Polymerase Chain ReactionABSTRACT
PCR implication using the primers for gag, pol and rev genes in BLV (bovine leukemia virus) proviral DNA and syncytium assay were carried out for the Korean native goats experimentally infected with bovine leukemia virus to investigate pathogenesis of BLV in the goats, and to establish a model animal for BLV infection. The oligonucleotide primers used in PCR revealed very high specificity, The minimal amount of FLK-BLV cellular chromosomal DNA to detect the integrated BLV proviral DNA was 10 ng. The peripheral blood lymphocytes from the goat infected with BLV were examined at regular intervals by PCR amplification and syncytium assay. Pol or gag genes were detected in none of three infected goats at the 1st week post-infection (p.i.). At the 4th week p.i., one of three goats showed the amplified gag gene. Thereafter detection rates for the genes were increased, indicating that the BLV proviral genes were integrated in all of the lymphocytes from three goats, at the 16th weeks p.i., when it was evident in syncytium assay that the lymphocytes from all of three goats were infested with infective BLV. Investigating the tissues from the necropsied goats at the 8th month p.i., the amplified BLV proviral genes and infective BLV were detected in all of the peripheral lymphocytes from three infected-goats. Among various tissues examined, the amplified BLV proviral genes were observed in spleen and superficial cervical, mandibular and retropharyngeal lymph nodes, and the infective BLV, in superficial cervical and mandibular lymph nodes. It was assumed that the Korean native goat was quite susceptible to BLV infection, indicating that the goat could be a good model animal for BLV.