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Detection of hepatitis C virus with RNA polymerase chain reaction in fulminant hepatic failure.

The role of hepatitis C virus (HCV) infection in fulminant hepatic failure is controversial. The frequency of serum HCV RNA positivity in previously reported patients with fulminant hepatic failure (FHF) of indeterminate cause ranged from 0 to 12% in the United States and Europe and from 43% to 59% in Asia. We assessed serum HCV RNA using polymerase chain reaction (PCR) and oligoprimers from the 5'UTR of the HCV genome in 26 consecutive patients with FHF. Another laboratory independently performed PCR on 21 of the serum samples using different oligoprimers from the 5'UTR and NS3 region of the …

Analysis of enterovirus genotypes using single-strand conformation polymorphisms of polymerase chain reaction products.

Enterovirus genotypes were identified rapidly by reverse (RT-PCR) followed by single-strand conformation polymorphism (SSCP) analysis. The primer pair was chosen from the highly conserved sequence at the 5' non-coding region of enterovirus genomes. RT-PCR amplified a 154 bp sequence in all samples from 14 serotypes of enteroviruses, including group A and B Coxsackie viruses, echoviruses and polioviruses. SSCP analysis of these products revealed different electrophoretic profiles. Thus, SSCP analysis will be useful for differentiating the genotypes of enteroviruses, and may be applicable for ra

Search for the human papillomavirus in nasal polyps, using a polymerase chain reaction-method.

Viral etiology of nasal polyps was postulated as many as 40 years ago, but so far, no study has shown an association or causal relation between any specific virus and nasal polyps. By using the polymerase chain reaction (PCR) technique, nasal polyps from both 10 patients with intolerance to nonsteroidal anti-inflammatory drugs (NSAID intolerance) (e.g., Aspirin) and from 10 patients with no history of NSAID intolerance were studied for the presence of papillomavirus DNA. Nasal mucosa from 10 patients who had undergone surgery for septoplasty served as controls. The PCR test used covers approxi…

Обнаружение ДНК верицелла зостера (VZV) в тканях плода с помощью цепной реакции полимеразы.

Первичная вирусная инфекция Варичеллы (VZV) во время беременности может быть связана с тяжелыми повреждениями плода. Однако вирологический диагноз врожденного синдрома VZV пока не увенчался успехом, поскольку VZV не мог быть изолирован от ткани плода. В настоящее время мы применили цепную реакцию полимеразы (ПХД), с тем чтобы можно было обнаружить ДНК VZV у плода, чьи матери приобрели ветрянку в первом триместре беременности. В 2 из 3 исследованных зародышей вирусная ДНК была обнаружена ПЦР независимо в 2 лабораториях, что подтверждает наличие прямой инфекции VZV плода.

Совпадение в аннотации оригинала
Primary Varicella Zoster Virus (VZV) infection during pregnancy can be associated with severe fetal malformation. The virological diagnosis of congenital VZV syndrome has not yet been successful, however, as VZV could not be isolated from fetal tissue. We have now applied a polymerase chain reaction (PCR) assay to allow the detection of VZV DNA in fetuses wh…

Detection of active cytomegalovirus infection in inflammatory aortic aneurysms with RNA polymerase chain reaction.

PURPOSE: We previously reported the possible role of human cytomegalovirus in the pathogenesis of inflammatory aortic diseases. To further analyze the viral cause of human aortic diseases, in this study we examined the presence and the replication of human Herpesviridae in 60 aortic tissues, including 7 inflammatory aneurysms, 37 atherosclerotic aneurysms, and 16 normal aortas. METHODS: To detect the genome of herpes simplex virus (type 1, type 2), cytomegalovirus, and Epstein-Barr virus, DNA polymerase chain reaction for each virus was performed. To analyze these herpesviral replications, th…

Polymerase chain reaction for detection of Borrelia coriaceae, putative agent of epizootic bovine abortion.

The nucleotide sequence of a chromosomally encoded antigen-expressing gene of Borrelia coriaceae was determined and used as a target for the polymerase chain reaction (PCR). Two primer sets were designed specifying the amplification of 269- and 701-bp DNA fragments. Primer set I, producing the short amplicon, was tenfold more sensitive than primer set II. As little as 10 fg of purified B coriaceae DNA could consistently be detected. The PCR assays, containing controlled numbers of whole spirochetes, allowed detectable amplification of 2 to 10 organisms. An internal, nonradioactively labeled ge…

[Development of a test system for detecting Leptospira interrogans using the polymerase chain reaction].

Based on polymerase chain reaction a test-system has been elaborated permitting one to identify the leptospirae of the most common serogroups (Icterohaemorrhagiae, Canicola, Javanica, Ballum, Pyrogenes, Pomona, Habdomadis, Sejroe, Tarassovi) of the species Leptospira interrogans. Sensitivity of the technique is 1-10 cells in a sample. The specificity of the system has been shown to depend on the temperature of the primers annealing. The elaborated system exceeds all other systems for leptospiral identification in sensitivity. It is prospective for leptospiral identification in biological liqui…

Detection of Chlamydia pneumoniae by polymerase chain reaction-enzyme immunoassay in an immunocompromised population.

Chlamydia pneumoniae has now been associated with pneumonia, bronchitis, pharyngitis, acute chest syndrome of sickle cell disease, and asthma. Because of the difficulty of primary isolation and tissue-culture adaptation of this organism, we used a previously developed polymerase chain reaction-enzyme immunoassay (PCR-EIA) to screen 132 culture-negative bronchoalveolar lavage (BAL) specimens from 108 immunocompromised patients (34% of whom were positive for human immunodeficiency virus) and 7 healthy volunteers. Thirteen specimens (9.8%) from 12 immunocompromised patients (11.1%) gave a positiv…

Unsuccessful attempt to detect genetic mutation in tuberous sclerosis utilizing the polymerase chain reaction.

Although tuberous sclerosis is supposed to be a phacomatosis inherited as an autosomal dominant trait, many cases develop without any affected parents or grandparents. In recent years, many vigorous investigations have been concentrated on finding the mutant gene, and possible candidate genes have been mapped on 9q34 and other chromosomes. In order to find a way of diagnosing asymptomatic carriers or patients, we tried to detect restriction fragment length polymorphisms (RFLPs) using the technique of polymerase chain reaction (PCR). We used a probe, MCOA12, which is located on 9q34 and has bee…

Polymerase chain reaction amplification of DNA from archival celloidin-embedded human temporal bone sections.

A method was developed for fast and efficient isolation of DNA from formalin-fixed, decalcified, celloidin-embedded human temporal bone sections for subsequent use in polymerase chain reaction (PCR) DNA amplification. The method relies on the use of an enzymatic digestion with proteinase K to release and solubilize the patient's DNA from an individual 20- to 25-microns temporal bone section. The method described should be of great value to those investigators extracting DNA from archival individual human temporal bone sections for polymerase chain reaction assays of specific genetic alteration…

Nested polymerase chain reaction for high-sensitivity detection of enteroviral RNA in biological samples.

A method based on nested polymerase chain reaction was developed for the detection of enteroviral genomes in biological samples. By taking advantage of the conserved 5' noncoding region of the enteroviral RNA, two sets of primers were utilized, enabling the detection either of a broad range of enteroviruses or of group B coxsackieviruses only. The sensitivity of the method is close to the detection of single molecules of viral RNA in as much as 1 mg of tissue sample. A preliminary study showed the usefulness of this technique for the analysis of endomyocardial biopsy samples from patients with…

Polymerase chain reaction and other methods to detect hot-spot and multiple gene mutations.

Gene mutations responsible for main genetic diseases as Duchenne/Becker muscular dystrophy or cystic fibrosis, and involved in more important diseases like cancer or cardiac diseases have been identified. Direct DNA tests can now be performed for these disorders. However, despite the knowledge of the exact alteration of the DNA sequence in these diseases, incorporating DNA analysis into large screening programs has been hindered by technical difficulties since each different mutation requires a different probe to be detected. To overcome these problems the polymerase chain reaction technique (…

Analysis of borderline-resistant strains of methicillin-resistant Staphylococcus aureus using polymerase chain reaction.

Identification of methicillin-resistant Staphylococcus aureus by drug-susceptibility tests alone poses a serious problem, because a considerable number of clinical S. aureus isolates are borderline resistant to methicillin. To circumvent this problem, we have developed a quick and sensitive method of PCR amplification for the detection of mecA gene, which, coding for PBP2', is the specific genetic element of methicillin-resistant Staphylococcus aureus. This method made it possible to identify MRSA strains in a short time using as few as 30 cells as a starting material for template DNA. Using t

Prenatal diagnosis of intrauterine infection with parvovirus B19 by the polymerase chain reaction technique.

Human parvovirus B19 is a recently recognized cause of fetal hydrops and death. Efforts to characterize the natural history of fetal infection with this virus have been hampered by the lack of sensitive and specific tests for diagnosis in utero. Using the highly sensitive polymerase chain reaction (PCR) assay, we determined the fetal infection status in 56 pregnancies by testing amniotic fluid, fetal serum, and maternal serum for B19 DNA and antibodies. Factors associated with a high risk of B19 infection were fetal disease, exposure to persons with erythema infectiosum, or signs or symptoms o…

Detection of pathogenic Entamoeba histolytica DNA in liver abscess fluid by polymerase chain reaction.

A sensitive method for detection of pathogenic Entamoeba histolytica DNA in drained fluids from liver abscess patients, using the polymerase chain reaction (PCR), has been developed. The PCR employs oligonucleotide primers specific for the gene encoding the 30 kDa molecule of pathogenic E. histolytica. Liver abscess fluids (19 samples), from 14 patients with a presumptive amebic liver abscess, were examined microscopically and by the PCR method. Only two of the 19 samples were positive microscopically, whereas all 19 samples tested positive by PCR. This technique can be used to confirm the dia…

Early detection of leukaemic relapse after bone marrow transplantation using the polymerase chain reaction.

We report a case of acute lymphoblastic leukaemia relapsing after allogeneic bone marrow transplantation in which the polymerase chain reaction (PCR) was used to assess chimeric status. This technique demonstrated the progressive reappearance of host cells prior to clinical relapse. The relapse was of host cell origin as shown by the presence of female (recipient) metaphases containing an abnormal chromosomal marker (iso 9q) which had also been present at initial diagnosis. The emergence of host cells in this case, detected only by PCR techniques but not by cytogenetic methods, appeared to her…

Polymerase chain reaction for detection of human cytomegalovirus infection in a blood donor population.

We have used the polymerase chain reaction (PCR) to analyse 420 normal donor blood samples taken at a city centre donation site. Three sets of human cytomegalovirus (HCMV) primers specific to the HXLF6, immediate early and late antigen gp64 genes, of the alpha, beta and gamma antigen coding regions respectively, were used to allow for the possibility of sequence variation. There was perfect correlation between the three sets of primers. Latex agglutination and enzyme-linked immunosorbent assay (ELISA) were also employed to provide data for a comparative study. The complete data show that infec…

Identification of mixed HIV-1/HIV-2 infections in Brazil by polymerase chain reaction.

Analysis of sera from hospitalized Brazilian patients by whole-virus lysate-based enzyme immunoassay and Western blot indicated that 0.4% were reactive to HIV-2 alone while 4% were reactive to both HIV-1 and HIV-2. When these sera were tested for HIV antibody by type-specific peptide enzyme immunoassays, dual seropositivity was confirmed in only 0.4% of patients. To define genetically the HIV strains within the population, we analyzed peripheral blood mononuclear cells from selected seropositive patients for the presence of HIV-1 and HIV-2 proviral DNA using the polymerase chain reaction (PCR)…

Detection of genital human papillomaviruses by polymerase chain reaction amplification with degenerate nested primers.

Degenerate oligonucleotide primers were designed for in vitro amplification by polymerase chain reaction (PCR) of a relatively well-conserved portion of the L1 capsid protein gene of genital human papillomaviruses. A specific 441bp fragment was amplified by PCR from genomic clones and clinical biopsy specimens containing DNA from HPV types 6, 11, 16, 18, 31, and 33, as well as from a number of other clinical specimens known to contain unclassified HPV isolates. As some HPV non-specific DNA was also often amplified, another set of degenerate primers was designed which amplified a 335 bp sequenc…