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Biomedical subjects

Yi-Wei Tang

Publications and source records attributed to Yi-Wei Tang.

At least 37 records · Page 2Linked to original sources

Dual infections of the central nervous system with Epstein-Barr virus.

We describe clinical and laboratory characteristics of 16 patients with central nervous system (CNS) infection caused by Epstein-Barr virus (EBV) and another pathogen. Seven of 10 immunocompromised patients had coinfection with viruses (3 with cytomegalovirus, 2 with JC virus, and 2 with varicella zoster virus) and 3 with nonviral pathogens (2 with pneumococcus and 1 with Cryptococcus species). Three of 6 immunocompetent patients had coinfections with viruses (1 each with herpes simplex virus, varicella zoster virus, and West Nile virus), and 3 had coinfections with nonviral pathogens (2 with Ehrlichia chaffeensis and 1 with Mycoplasma pneumoniae). The EBV load was similar in immunocompromised and immunocompetent patients and in patients with viral and nonviral coinfections. EBV lytic-cycle mRNA was detected in the cerebrospinal fluid of 5 of 6 tested samples, indicating EBV replication in the CNS during coinfection.

Adult↗

Detection of hepatitis C virus by a user-developed reverse transcriptase-PCR and use of amplification products for subsequent genotyping.

BACKGROUND: Detection, quantitation and genotyping of hepatitis C virus (HCV) are important in selecting appropriate therapy. Current commercially available HCV genotyping kits, including sequencing-based TRUGENE HCV 5'NC and hybridization-based INNO-LiPA HCV II assays, rely on amplification products (amplicons) generated by HCV reverse-transcriptase (RT)-PCR methods such as the Roche AMPLICOR HCV test. METHODS: We developed a one-step RT-PCR assay to amplify and detect HCV RNA, and the resulting amplicons were used for HCV genotyping (TRUGENE). A total of 142 clinical samples were used to compare results from the RT-PCR/TRUGENE assay and those generated by the COBAS AMPLICOR and INNO-LiPA tests. RESULTS: Eighty-seven of 108 plasma specimens which were positive by AMPLICOR were also positive by the user-developed RT-PCR, giving a sensitivity of 100.0%. The RT-PCR detected 2 of 21 AMPLICOR-negative specimens and none of 34 HCV-EIA-negative serum specimens, giving a specificity of 96.4%. The 87 amplicons from the RT-PCR yielded HCV genotypes. HCV genotype results from both TRUGENE and INNO-LiPA were all in agreement. The TRUGENE and INNO-LiPA assays identified 69 (79.3%) and 47 (54.0%) specimens, respectively at the subtype level. HCV subtype information agreed by both assays in 34 of 36 (99.4%) specimens. One specimen with HCV genotypes 2 and 4 by INNO-LiPA was classified as a single genotype 2 by TRUGENE. CONCLUSION: Our data showed that the user-developed RT-PCR has comparable sensitivity and specificity for the detection of HCV in clinical specimens. The amplicons generated by the RT-PCR can be used for HCV genotyping by the sequencing-based TRUGENE assay.

Base Sequence↗

Cytokine pattern is solely influenced by priming vaccine but immunity and disease by both priming and boosting vaccines in mice challenged with respiratory syncytial virus.

Vaccine formulation can influence cytokine and disease patterns in mice following respiratory syncytial virus (RSV) challenge. The influence of different live and killed dual-vaccine combinations on subsequent immune responses was investigated. BALB/c mice received either killed followed by killed (KV/KV), killed followed by live (KV/LV), live followed by killed (LV/KV), or live followed by live (LV/LV) RSV vaccines intramuscularly. Mouse weight loss, viral replication, cytokine expression patterns, immunoglobulin isotype antibody profiles, neutralizing antibody responses, and cytotoxicity T lymphocyte (CTL) activities in lungs were compared on subsequent live RSV challenge. On challenge, mice vaccinated initially with KV and boosted with either KV or LV expressed significantly skewed ratios of IL-4 to IFN-gamma mRNA and IgG1 to IgG2a antibody, when compared to those vaccinated initially with LV. Low levels of RSV replication were detected in lungs of mice vaccinated with KV/KV, KV/LV, and LV/KV, but not in mice vaccinated with LV/LV. Mice vaccinated with KV/LV, LV/KV, or LV/LV had RSV-specific CTL activity in lungs six days after RSV challenge, while no CTL activity was detected in KV/KV-vaccinated mice. Mice vaccinated with KV/KV had the greatest weight loss, while LV/LV-vaccinated mice resulted in the least. Mice vaccinated with either KV/LV or LV/KV had intermediate weight loss after challenge. These data indicate that an original antigenic sin-like phenomenon was exhibited in cytokine and immunoglobulin isotype responses in mice after challenge. T helper (Th)-like immune responses were determined solely by the initial vaccination, while weight loss, viral replication, neutralizing antibody responses, and CTL activities were also influenced by boosted vaccinations.

Animals↗

Normal values for CD4 and CD8 lymphocyte subsets in healthy Chinese adults from Shanghai.

The aim of this study was to establish reference ranges for lymphocyte subsets in Chinese adults. Venous blood specimens were obtained from 614 healthy, human immunodeficiency virus (HIV)-seronegative adults in Shanghai. Flow cytometry was used to determine percentages and absolute numbers of CD4 and CD8 T lymphocytes. Mean values for CD4 and CD8 lymphocytes were 727 and 540 cells/microl, respectively, yielding a CD4/CD8 ratio of 1.49. While CD8 lymphocyte values varied with age and gender, no significant differences in CD4 lymphocyte values were observed. Shanghai adults had approximately 100 fewer CD4 lymphocytes/microl on average than Caucasians, suggesting that lower CD4 lymphocyte cutoffs for classifying and monitoring HIV infection may be needed in China.

Adolescent↗

Surveillance of childhood influenza virus infection: what is the best diagnostic method to use for archival samples?

Despite the clinical importance of influenza virus in pediatric respiratory infections, the optimal set of diagnostic tests to use when conducting studies using archival samples is not clear. In this study, we compared diagnostic tests for influenza virus in 75 children younger than 5 years of age who presented with symptomatic respiratory infection during one of four influenza seasons, had negative viral cultures for other respiratory pathogens, and had both an archival nasal aspirate obtained at the time of illness and serology spanning that influenza season. For all eligible children, we compared the results of viral culture performed at the time of collection with serology and PCR of archival nasal aspirates. Using real-time viral culture as the "gold standard," the test characteristics of PCR of archival nasal aspirates (sensitivity, 82%; specificity, 100%) and serology (sensitivity, 82%; specificity, 87%) were similar. The relatively low sensitivity of PCR of archival nasal samples in this study compared to that of PCR of fresh samples in a previous study suggests that RNA degradation occurred despite storage of the specimens at -70 degrees C. RNA degradation would also explain why only 11 (52%) of 21 archival nasal samples that had positive influenza virus cultures at the time of collection had positive repeat cultures in the summer of 2000. Thus, in archival specimens stored at -70 degrees C, PCR was more sensitive than viral culture. However, testing of fresh specimens had the highest yield in this study. Studies of optimal methods for specimen storage are needed.

Child, Preschool↗

Identification of the bacterial etiology of culture-negative endocarditis by amplification and sequencing of a small ribosomal RNA gene.

We report two cases of culture-negative bacterial endocarditis in which the organisms were identified by amplification and sequencing of the bacterial 16S rRNA gene. These results support an important role for polymerase chain reaction followed by direct sequencing to determine the etiology of culture-negative bacterial endocarditis and to guide appropriate antimicrobial therapy.

Adult↗

Measurement of human cytomegalovirus loads by quantitative real-time PCR for monitoring clinical intervention in transplant recipients.

Quantitative monitoring of human cytomegalovirus (HCMV) infection is helpful in determining appropriate antiviral management of transplant recipients. Quantitative PCR technologies have demonstrated accuracy in measuring systemic HCMV loads. A total of 298 consecutive whole-blood specimens submitted to the Clinical Virology Laboratory at Vanderbilt University Medical Center from 15 February to 31 October 1999 were included in the study. In addition to a qualitative colorimetric microtiter plate PCR assay (MTP-PCR) and a semiquantitative pp65 antigenemia assay, each specimen was measured for HCMV loads by a quantitative PCR assay performed on an ABI PRISM 7700 Sequence Detection System (TaqMan). Compared to results of the MTP-PCR, the sensitivity, specificity, positive predictive value, and negative predictive value were 70.5, 97.5, 87.8, and 92.8% for the antigenemia assay and were 96.7, 92.0, 75.6, and 99.1% for the TaqMan assay, respectively. There was a high correlation between antigenemia values and HCMV loads as determined by the TaqMan (r = 0.989; P < 0.001). Antigenemia values of 0, 1 to 10, 11 to 100, 101 to 1,000, and over 1,000 positive cells per 2 x 10(5) leukocytes corresponded to median HCMV loads measured by TaqMan of 125, 1,593, 5,713, 16,825, and 5,425,000 copies/ml, respectively. Corresponding to antigenemia values of 1 to 2, 10, and 50 positive cells per 2 x 10(5) leukocytes, HCMV viral loads of 1,000, 4,000, and 10,000 copies/ml are proposed as cutoff points for initiating antiviral therapy in patient groups with high, intermediate, and low risk of CMV diseases.

Cytomegalovirus↗

Characterization of a novel rapidly growing Mycobacterium species associated with sepsis.

A rapidly growing mycobacterium was isolated five times from blood cultures from a 6-year-old female patient with relapsed pre-B-cell acute lymphocytic leukemia. All five isolates had identical nucleotide sequences for the first 500 bp of the 16S rRNA gene, indicative of a single species. High-performance liquid chromatography analysis of mycolic acids indicated that the species was similar to Mycobacterium smegmatis. Sequence analysis of the 16S rRNA gene (1,455 bp) for one isolate demonstrated that the species was closely related to Mycobacterium diernhoferi. Based on the phenotypic features and phylogenetic analysis, it was concluded that the isolates represented a novel rapidly growing Mycobacterium species. The name "Mycobacterium hackensackense" is proposed for this unique strain, 147-0552(T), which was deposited in the American Type Culture Collection as ATCC BAA-823(T).

Base Sequence↗

Genetic analysis of Crimean-Congo hemorrhagic fever virus in Russia.

Genetic analysis of wild-type Crimean-Congo hemorrhagic fever (CCHF) virus strains recovered in the European part of Russia was performed. Reverse transcriptase PCR followed by direct sequencing was used to recover partial sequences of the CCHF virus medium (M) genome segment (M segment) from four pools of Hyalomma marginatum ticks and six human patients. Phylogenetic analysis of the M-segment sequences from Russian strains revealed a close relatedness of the strains (nucleotide sequence diversity, <or=5.0%). The strains differed significantly from CCHF viruses from other regions of the world (nucleotide sequence diversity, 10.3 to 20.4%), suggesting that CCHF virus strains recovered in the European part of Russia form a distinct group.

Genome, Viral↗

Herpesvirus DNA is consistently detected in lungs of patients with idiopathic pulmonary fibrosis.

On the basis of earlier reports associating Epstein-Barr Virus (EBV) with half of the cases of idiopathic pulmonary fibrosis (IPF), we hypothesized that chronic infection with EBV or a closely related herpesvirus would be detected in all cases of IPF. We tested lung specimens from 33 IPF patients (8 patients with familial IPF and 25 patients with sporadic IPF) and 25 patients with other diseases as controls for the presence of eight herpesviruses using PCR-based techniques. One or more of four herpesviruses (cytomegalovirus [CMV], EBV, human herpesvirus 7 [HHV-7], and HHV-8) were detected in 32 of 33 (97%) subjects with IPF and in 9 of 25 (36%) controls (P < 0.0001). CMV, EBV, and HHV-8 were found more frequently in IPF patients than in controls (P < 0.05, P < 0.001, and P < 0.01 respectively). Two or more herpesviruses were detected in 19 of 33 (57%) IPF patients and in 2 of 25 (8%) controls (P < 0.001). Two or more herpesviruses and HHV-8 were found more frequently in patients with sporadic IPF than in patients with familial IPF (P < 0.05 for both comparisons), and CMV was found less frequently in patients with sporadic IPF than in patients with familial IPF (P < 0.05). Immunohistochemistry for EBV or HHV-8 antigen showed viral antigen primarily in airway epithelial cells. These data support the concept that a herpesvirus could be a source of chronic antigenic stimulation in IPF.

Adult↗

Major outbreak of toxic shock-like syndrome caused by Streptococcus mitis.

Severe illness caused by viridans streptococci rarely occurs in immunocompetent hosts. Between December 1990 and May 1991, thousands of patients in the YangZi River Delta area of Jiangsu Province, China, suffered from scarlet fever-like pharyngitis. Fewer cases occurred in subsequent years with the same seasonality. Approximately half of the cases developed complications characteristic of streptococcal toxic shock-like syndrome (TSLS). Throat cultures yielded predominant growth of alpha-hemolytic streptococci. All cases admitted to Haian People's Hospital were investigated. Clinical specimens were collected, medical records were reviewed, and bacterial isolates were identified phenotypically and analyzed by 16S rRNA gene sequencing and pulsed-field gel electrophoresis (PFGE). Proteins were purified from culture supernatants by extraction, ammonium sulfate precipitation, and fast-protein liquid chromatography. Biological activities of protein components were determined by subcutaneous inoculation into rabbits. A total of 178 cases of non-beta-hemolytic streptococcal scarlet fever-like pharyngitis were studied. In 88 (79.3%) of 111 patients, oropharyngeal swab cultures grew morphologically identical alpha-hemolytic streptococci. A protein in culture supernatants was pyrogenic in rabbits, was mitogenic for splenocytes, and enhanced rabbit susceptibility to endotoxin challenge. The N-terminal amino acid sequence of this 34-kDa protein showed no homology with known Streptococcus pyrogenic exotoxins. The organism was identified as Streptococcus mitis based on biochemical and 16S rRNA sequence analyses. Representative outbreak isolates from 1990 to 1995 displayed identical PFGE patterns. This TSLS outbreak in southeastern China was caused by a toxigenic clone of S. mitis. An apparently novel toxin may explain the unusual virulence of this organism.

Animals↗

Molecular diagnostics of atypical pneumonia.

The emergence of nucleic acid-based molecular techniques has significantly enhanced laboratory diagnosis and monitoring of atypical pneumonia. These techniques have not only provided rapid and sensitive detection of fastidious microbial organisms but have also played critical roles in identifying and characterizing emerging pathogens that cause atypical pneumonia. Other benefits that molecular techniques can bring to the field include organism differentiation, quantitation, typing, and antibiotic resistance profiles. Gradually becoming standardized and widely available, the future will see some promising molecular methods become a mainstay in clinical laboratories for recognition and diagnosis of atypical pneumonia pathogens.

Chlamydia↗

Molecular Techniques in the Diagnosis of Central Nervous System Infections.

Development of polymerase chain reaction (PCR)-based molecular techniques has initiated a revolution in the field of diagnostic microbiology. These techniques have not only provided rapid, noninvasive detection of microorganisms that cause central nervous system (CNS) infections, but have also demonstrated that several neurologic disorders are linked to infectious agents. While PCR-based techniques are predicted to be widely used in diagnosing and monitoring CNS infections, the limitations, as well as strengths, of these techniques must be clearly understood by both clinicians and laboratory personnel to ensure proper utilization.

Journal Article↗

Prevalence of methicillin-resistant Staphylococcus aureus nasal carriage in the community pediatric population.

BACKGROUND: Reports from various geographic regions indicate that the prevalence of community-acquired methicillin-resistant (MRSA) infection is increasing. The primary reservoir is the anterior nares; nasal carriage is a risk factor for infection in a variety of populations. Little is known about MRSA nasal carriage rates among children in Nashville, TN and the associated likelihood of community MRSA transmission. METHODS: Nasal swabs were collected from 500 children at well-child visits at either a university hospital pediatric clinic or a private pediatric office. Cultures were plated onto selective staphylococcal media, with or without oxacillin. isolates were confirmed by coagulase tube testing. Antibiotic susceptibilities were determined for suspected methicillin-resistant isolates by standard broth microdilution methods (National Committee for Clinical Laboratory Standards). Pulsed field gel electrophoresis was used to evaluate epidemiologic relatedness. PCR testing was done to assess for the gene. A parent questionnaire was administered regarding MRSA risk factors. RESULTS: Four patients had oxacillin-resistant isolates (MIC >or= 4 microg/ml), and two had borderline resistant isolates (MICs = 1 and 2 microg/ml). One of the borderline-resistant isolates and one of the MRSA isolates had pulsed field gel electrophoresis typing results indicating close relatedness. The gene was present in all resistant isolates and one of the borderline-resistant isolates. Only having a household member employed in a hospital was associated with a greater risk of MRSA nasal carriage (odds ratio, 9.6; P= 0.008). CONCLUSIONS: MRSA nasal colonization is present within Nashville's healthy pediatric population. Children with household contacts employed in a hospital are significantly more likely to be colonized.

Anti-Bacterial Agents↗

Rapidly increasing prevalence of penicillin-resistant Streptococcus pneumoniae in middle Tennessee: a 10-year clinical and molecular analysis.

The clinical and molecular epidemiology of penicillin-resistant Streptococcus pneumoniae and the diagnostic accuracy of a six-primer PCR assay in identifying penicillin resistance were analyzed by using clinical isolates recovered over a 10-year period in middle Tennessee. The prevalence of non-penicillin-susceptible S. pneumoniae isolates (MIC, > or =0.1 microg/ml) increased from 10% in 1990 to 70% in 1999 (P < 0.001). Among S. pneumoniae isolates for which the penicillin MIC was > or =2 microg/ml (highly penicillin-resistant S. pneumoniae [PRSP]), 23 and 5% were resistant to at least three and at least five other antimicrobial classes, respectively. Pulsed-field gel electrophoresis identified 13 unique strain types, with type B accounting for 33% of PRSP isolates. The sensitivity, specificity, and negative and positive predictive values of the PCR assay in detecting PRSP were 99, 100, 99, and 100%, respectively. Penicillin resistance is rapidly increasing among S. pneumoniae isolates in Tennessee. The simultaneous detection of S. pneumoniae and high-level penicillin resistance can be accurately performed with the six-primer PCR assay.

Adolescent↗

Enterococcus faecium-related outbreak with molecular evidence of transmission from pigs to humans.

Between 24 July and 31 August 1998, thousands of domestic pigs died of hemorrhagic shock in three adjunct counties along the YangZi River in Jiangshu Province, China. From 28 July to 6 September 1998, 40 local farmers (36 males and 4 females, ages 23 to 78 years) were hospitalized with severe illness characterized by high fever, erythematous rash or petechiae, and profound lethargy after contact with sick pigs. Twelve (30%) of these patients died of respiratory failure and shock. Eleven bacterial isolates recovered from 11 blood and cerebrospinal fluid specimens collected from seven patients and two pigs were identified as Enterococcus faecium based on biochemical reactions and 16S rRNA gene sequence analysis. Both pig and human E. faecium isolates displayed indistinguishable antibiotic susceptibility and pulsed-field gel electrophoresis patterns. These data strongly suggest the spread of an outbreak of E. faecium-related sepsis from pigs to humans.

Adult↗