Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0149514 (bronchitis)
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Polymerase chain reaction (PCR) using primers targeting the IS6110 repetitive sequence was employed to detect Mycobacterium tuberculosis in 228 samples from patients with tuberculosis or other pulmonary diseases and controls, and the results were compared with culture and clinical findings. None of culture negative samples from 17 healthy controls were PCR positive. Of 109 active tuberculosis patients under chemotherapy, 88 (80.7%) were PCR positive and were significantly higher than 63 (57.8%) positive by culture. Fifty-nine (93.7) of 63 culture positive and 29 (63.0%) of 46 culture negative specimens contained M. tuberculosis detectable by PCR. In 41 specimens from inactive tuberculosis patients who visited to the chest clinic because of chest problems, 16 (39.0%) also gave PCR positive results. In addition, 14 (46.7%) of 30 specimens submitted for M. tuberculosis culture from patients with pulmonary diseases were PCR positive. Presumptive diagnosis of these PCR positive patients was bronchitis, pneumonia, bronchial asthma, etc. Therefore, this study suggests that PCR is sensitive and specific in detecting M. tuberculosis in clinical specimens. However, the interpretation of the PCR results in specimens from patients with pulmonary diseases should be done cautiously in areas with a high prevalence of tuberculosis.
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PMID:Detection of Mycobacterium tuberculosis in clinical samples from patients with tuberculosis or other pulmonary diseases by polymerase chain reaction. 129 44

A workshop in which 17 practicing scientists participated was intended to address primarily people who use or could use biotechnology in their work and was confined to five techniques. Endonuclease fingerprinting and mapping involved cleaving nucleic acid with a specific restriction enzyme and separating the nucleic acid fragments by electrophoresis. Field and vaccine isolates of Pasteurella multocida could be distinguished; Salmonella enteritidis could be divided into three groups; chlamydia could be grouped into seven groups; and vaccinia, quail pox, and fowl pox could be clearly distinguished. Preparation of nucleic acid probes involved producing large amounts of labeled oligonucleotides, usually of unknown sequence. Successful probes had been made for infectious bursal disease virus, avian influenza virus, Newcastle disease virus, and infectious bronchitis virus. In Southern, Northern, and dot blotting, either DNA or RNA fragments were placed on or transferred to a solid substrate and probed. The procedure was able to detect infectious bursal disease virus, infectious bronchitis virus, Mycoplasma gallisepticum, and Marek's disease virus. In situ hybridization involved applying a labeled probe to frozen or fixed sections or to intact cells. In Polymerase chain reaction, two primers, some distance apart, were annealed to a denatured target DNA. Repeated cycles of DNA synthesis with a thermostable polymerase, denaturing, and reannealing resulted in great amplification of a rare sequence. After 30 cycles, a rare gene sequence could be amplified more than 10(6) times. It was used successfully to detect minute quantities of influenza virus and infectious bursal disease virus, and the process was used to facilitate DNA sequencing of coccidiosis gene segments.
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PMID:Practical application of nucleic acid techniques to avian disease problems. 255 97

This recently recognised member of the genus Chlamydia is one of the most widespread pathogens of man, though up to 90% of infected people have few or no symptoms. Several studies have estimated the population prevalence of antibodies to C. pneumoniae at 40-55% in the northern hemisphere, and over 60% in under-developed countries. The incidence of infections follows a cyclical pattern, with peaks at regular intervals of 2-10 years, but no apparent seasonal periodicity. Nosocomial transmission may be mediated by environmental surfaces as well as aerosols, and immunosuppression, for example by the human immunodeficiency virus, predisposes to infection. Chlamydia pneumoniae causes predominantly atypical pneumonia, often severe in adults, especially the elderly; including 5-10% of community-acquired pneumonia in Scandinavian countries. Serological evidence indicates associations with asthma, bronchitis, exacerbations of chronic airflow obstruction, otitis media and bronchiolitis. Several studies, using both serological and morbid anatomical techniques, also indicate associations with vascular atheroma and ischaemic heart disease, and with acute myocardial infarction. Chronic, latent and recurrent infections have been documented, and it is postulated that, like chronic or recurrent C. trachomatis infections, these may produce disease as a consequence of the host's immunological hypersensitivity. Several techniques are available for serological diagnosis: the technique of choice is micro-immunofluorescence, using fixed whole elementary or reticulate bodies as antigen, but antibody responses are highly variable. Traditional alternatives, antigen detection (by direct immunofluorescence or enzyme immunoassay) and cell culture, have major disadvantages. Polymerase chain reactions have not yet been widely applied to the clinical setting. tetracycline antibiotics, erythromycin and quinolones are not very efficacious in the treatment of C. pneumoniae infection. The azalide antibiotic, azithromycin, and the macrolide, clarithromycin, are active in vitro against C. pneumoniae, and may become treatments of choice. The development of anti-chlamydial vaccines remains an important research goal.
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PMID:Clinical aspects of Chlamydia pneumoniae infection. 789 84

Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis were used to differentiate between serotypes of several infectious bronchitis virus (IBV) strains. A sequence of 1720 base pairs (bp) that contains the S1 glycoprotein gene of IBV was amplified by PCR, purified, and digested with restriction enzymes. Eleven reference IBV strains were grouped according to the RFLP patterns. The IBV Holte, Arkansas DPI, SE 17, Md 27, and Iowa 97 strains could be differentiated from the other IBV strains using the restriction enzyme HaeIII. The Beaudette, Massachusetts 41, Connecticut, and Florida 88 strains had the same HaeIII RFLP pattern but could be differentiated using XcmI and BstYI restriction enzymes. The Gray and JMK strains could not be differentiated by their RFLP patterns following digestion with 23 different restriction enzymes. Twenty-six samples (field isolates and reference strains) of IBV, previously serotypes by the virus-neutralization (VN) test in embryonating eggs, were analyzed in a blind fashion. The results using the PCR and RFLP analysis agreed with the serotype for traditional and variant IBV viruses as determined by the VN test.
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PMID:Differentiation of infectious bronchitis virus serotypes using polymerase chain reaction and restriction fragment length polymorphism analysis. 809 82

Polymerase chain reaction (PCR) and a biotin-labeled DNA probe were used to amplify and detect the genome of infectious bronchitis virus (IBV) from tracheal swabs taken from chickens that were experimentally inoculated with the IBV Beaudette, Arkansas, and Gray strains. The viral genome was successfully detected by PCR and confirmed by dot-hybridization assay using a biotin-labeled DNA probe on days 1, 3, 9, and 14 after exposure. Direct electron microscopy (EM) analysis was used to compare the ability of the two tests to detect IBV from the same tracheal swab samples. The EM analysis did not detect IBV in four of eight necropsy groups that were positive using PCR and the biotin-labeled DNA probe. Although histopathological lesions were observed in the tracheas, no clinical signs or specific antibody response were observed in the birds. The virus was also detected in the allantoic fluid of embryonating chicken eggs that had been inoculated with field samples suspected to be IBV. The field samples were passed four to six times in embryonating eggs, and 10 of 17 samples were positive using PCR and the biotin-labeled probe.
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PMID:Polymerase chain reaction and a biotin-labeled DNA probe for detection of infectious bronchitis virus in chickens. 838 58

A 18-year-old male was admitted to another hospital complaining of his chest X-ray. After transfer to our hospital, increased serum antibody titers to simultaneous M. pneumoniae and C. psittaci were noted. These antibody titers decreased after about four months. Positive results for M. pneumoniae was obtained by Polymerase chain reaction in the right pleural effusion. Based on these findings, this case was diagnosed as M. pneumoniae and C. psittaci pneumonia. A transbronchial lung biopsy and a bronchial biopsy revealed rare histological findings, including histiocytic intra-alveolar pneumonia with palisaded granuloma and small foci of necrosis in the left upper lobe and eosinophilic bronchitis in the right middle bronchus. His chest X-ray and chest CT showed a nodular shadow, obstructive pneumonia and pleural effusion which are rare in M. pneumoniae and C., psittaci pneumonia.
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PMID:[An adult case of pneumonia due to Mycoplasma pneumoniae and Chlamydia psittaci]. 882 57

Enteroviruses are members of Enterovirus genus of Picornaviridae family. On the basis of their pathogenesis and host range, most human enteroviruses are classified into one of three groups (Coxsackie's viruses, echoviruses and polioviruses). Some unclassified human enteroviruses may cause bronchitis (type 68), acute hemorrhagic conjunctivitis (type 70), meningitis and paralysis (types 70 and 71) and hepatitis (type 72 or hepatitis A virus). Enteroviruses can be propagate in primary cultures of human monkey kidney cells and in some cell lines such as HeLa, Vero and WI-38. Virions are small (22-30 nm diameters) containing ss RNA, monopartite and have icosahedra symmetry. The fast, high sensitive and specific detection of enteroviruses today is achieved by using of PCR (Polymerase chain reaction) method. But, PCR is so much more than just mixing reagents in a tube and running a thermal cycler. In each laboratory with PCR facilities, it is necessary to find optimal PCR conditions (performing of PCR optimization experiments). In this paper we presented results of PCR optimization for enteroviruses by using of poliovirus type 1(Sabin). Optimal obtained PCR parameters were: 2,5 mM MgCl2, dNTPs dilution 10(-1) and annealing temperature 50 degrees C, after 30 amplification cycles in Perkin Elmer 2400 thermal cycler.
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PMID:Optimisation of RT-PCR for detection of enteroviruses. 1676 11

This study clinically and molecularly characterizes an adenovirus epidemic that broke out in Taiwan in April 2004. Clinical data on 325 children diagnosed with acute illness were collected between April 2004 and April 2005, and a diagnosis of adenovirus was confirmed by viral isolation. Polymerase chain reaction and restriction fragment length polymorphism were used to identify the adenovirus genotypes in 267 patients. There was a seasonal variation, with a peak incidence between November 2004 and January 2005 (p < 0.001). The median age was 52 months, range 1-210 months. Most cases (90.8%) were younger than 7 years old. Male-to-female ratio was 1.56:1. The most common clinical diagnosis was exudative tonsillitis (50.8%), followed by bronchitis/bronchiolitis (29.9%), conjunctivitis or pharyngoconjunctival fever (22.5%), and acute otitis media (16.3%). Adenovirus type 3 was found in 215 patients (80.5%). The other 52 patients had other genotypes: type 2 (10.1%), type 1 (6.0%), type 5 (1.9%), type 7 (0.7%), type 4 (0.4%), and type 6 (0.4%). Patients with type 3 were significantly older [age >52 months, adjusted odds ratio (OR) 8.55, 95% confidence interval (CI) 1.84-40, p = 0.006), their family members had a higher incidence of illness (adjusted OR 8.77, 95% CI 1.55-50, p = 0.01), they coughed (adjusted OR 6.37, 95% CI 1.54-26.3, p = 0.01), and they had a higher C-reactive protein (CRP) level (>2.87 mg/dL, adjusted OR 3.64, 95% CI 1.06-12.3, p = 0.04) than the 52 cases with other genotypes. In conclusion, this adenovirus outbreak, from late autumn to winter, was predominately caused by adenovirus type 3. Patients with this genotype were significantly older, had a higher incidence of cough and family transmission, and had higher CRP levels than those with other genotypes.
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PMID:Molecular and clinical characteristics of adenoviral infections in Taiwanese children in 2004-2005. 1787 5

The biological nature of vaccines imposes a permanent risk for contamination with extraneous agents. Therefore, testing of vaccines for freedom from extraneous agents is essential in the manufacturing process and quality control. Relevant methods for testing for extraneous agents of avian viral vaccines are specified in the monographs of the European Pharmacopoeia (Ph. Eur.). Currently, most of these methods involve the use of embryonated eggs or chickens. Polymerase chain reaction (PCR) is a widely used and suitable tool for the amplification and detection of extraneous nucleic acids. Different PCR assays have been developed for the application in routine testing of veterinary vaccines. However, before introduction of new methods in monographs of the Ph. Eur., they must undergo validation. Here we report about a pre-validation study performed in Official Medicines Control Laboratories (OMCLs). Diluted samples of avian infectious laryngotracheitis, avian infectious bronchitis and avian infectious bursal disease viruses have been analysed using standardised procedures and reagents. The study demonstrated that PCR methods can be transferred to other laboratories. The results also show that further work is warranted for full validation of the method.
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PMID:Pre-validation study for testing of avian viral vaccines for extraneous agents by PCR. 1841 34

Polymerase chain reaction (PCR)-based approaches to the detection, differentiation and characterization of avian pathogens continue to be developed and refined. The PCRs, or reverse transcriptase-PCRs, may be general, designed to detect all or most variants of a pathogen, or to be serotype, genotype or pathotype specific. Progress is being made with respect to making nucleic acid approaches more suitable for use in diagnostic laboratories. Robotic workstations are now available for extraction of nucleic acid from many samples in a short time, for routine diagnosis. Following general PCR, the DNA products are commonly analyzed by restriction endonuclease mapping (restriction fragment length polymorphism), using a small number of restriction endonucleases, based on a large body of sequence data. Increasingly, however, nucleotide sequencing is being used to analyze the DNA product, in part due to the expanding use of non-radioactive sequencing methods that are safe and enable high throughout. In this review, I highlight some recent developments with many avian viruses: Newcastle disease virus; circoviruses in canary and pigeon; infectious bursal disease virus (Gumboro disease virus); avian adenoviruses, including Angara disease/infectious hydropericardium virus, haemorrhagic enteritis virus of turkeys, and egg drop syndrome virus; avian herpesviruses, including infectious laryngotracheitis virus, duck plague virus, psittacine herpesvirus (Pacheco's parrot disease virus), Marek's disease virus and herpesvirus of turkeys; avian leukosis virus (associated with lymphoid leukosis or myeloid leukosis, and egg transmission); avian pneumoviruses (turkey rhinotracheitis virus); avian coronaviruses, including infectious bronchitis virus, turkey coronavirus and pheasant coronavirus; astrovirus, in the context of poult enteritis and mortality syndrome, and avian nephritis virus; and avian encephalomyelitis virus, a picornavirus related to hepatitis A virus.
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PMID:Innovation and discovery: the application of nucleic acid-based technology to avian virus detection and characterization. 1918 52


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