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[Polymerase chain reaction for rapid detection and serotyping of dengue virus in clinical samples].

This study describes the benefits of using reverse transcriptase polymerase chain reaction (RT-PCR) for the rapid detection and typing of dengue virus in clinical samples. Twenty-seven serum specimens from patients with dengue fever and dengue hemorrhagic fever in Colombia, Nicaragua, and Panama were directly subjected to RT-PCR for the detection of dengue virus. The resulting double-stranded DNA product was typed by a second round of PCR amplification (nested PCR) with type-specific primers, viral culture/indirect immunofluorescence (IIF), and enzyme-linked electroimmunoassay for IgM anti-den…

Diagnostic utility of the polymerase chain reaction in 2 cases of suspected Whipple disease.

We describe 2 patients with a diagnosis of Whipple disease in whom the usual antibiotic therapy failed. A polymerase chain reaction-based test was used to identify the recently described Whipple bacillus, Tropheryma whippelii. In one case, the diagnosis was confirmed, whereas in the second case, which had been histologically diagnosed as Whipple disease of the brain, the process was identified as a monocyte-derived histiocytosis. In conclusion, Whipple disease can be distinguished from other diseases with similar histological features with the use of a polymerase chain reaction-based test.

[Detection of circulating neoplastic cells by reverse transcriptase and polymerase chain reaction in melanoma].

BACKGROUND: Circulating melanocytes can be detected in peripheral blood of patients with malignant melanoma by means of tyrosinase messenger … d 2 à 8 weeks after. To identify circulating melanocytes, we used coupled reverse-transcription and polymerase chain reaction to target tyrosinase messenger RNA. RESULTS: None of the 20 patients with stage II melanoma had detectable circulating melanocytes before lymph node dissection. By contrast, 7 of them became transiently PCR positive in the 8 weeks following surgery …

A simplified polymerase chain reaction assay for detection of chromosomal translocations in hematologic malignancies.

The polymerase chain reaction (PCR) is a rapid and highly sensitive method for detection of a variety of chromosomal translocations in malignant tissues. Detection of each different type of translocation, or even DNA rearrangements at different breakpoint cluster regions within the same type of translocation, usually requires separate thermocycling parameters and/or buffer conditions. In this report, we describe a single set of reaction conditions, making use of progressively decreasing annealing temperatures and a standardized reaction buffer, that permits the detection of several different t…

Detection of minimal residual disease in leukemia using polymerase chain reaction and restriction analysis.

OBJECTIVE: To investigate the relationship between the clinical outcome of childhood acute lymphoblastic leukemia and the status of minimal residual disease after termination of therapy. METHODS: A total of 18 children with acute lymphoblastic leukemia were studied. Eleven cases were treated with chemotherapy, the other 7 with autologous peripheral blood stem cell transplantation (ABSCT) or autologous bone marrow transplantation (ABMT). The outcome of the 18 patients was obtained by clinical follow-up. Minimal residual disease was detected by means of the polymerase chain reaction (PCR) and r…

[Detection of human cytomegalovirus infection in blood donor population by the polymerase chain reaction].

Samples of peripheral blood from 76 different donors were detected for the presence of human cytomegalovirus (HCMV) DNA by polymerase chain reaction (PCR). The results showed that HCMVs were present in 39 out of 76 blood donors which reached an infection positive rate of 51.3%. In addition, the sensitivity and specificity on PCR were tested, it also indicated that the sensitivity can reach the level of 5fg.microliter-1. The method is simple, rapid, highly sensitive and specificity. It can be considered as a diagnostic measure in clinical detection of HCMV.

Detection of Chlamydia trachomatis in urine samples by polymerase chain reaction and enzyme immunoassay.

First-void urine samples collected from sexually transmitted diseases (STD) clinic patients were examined by a nested polymerase chain reaction (PCR) and a commercial enzyme immunoassay (IDEIA Chlamydia) for the diagnosis of Chlamydia trachomatis urethritis or cervicitis. The primers for the PCR amplified a target in the major outer membrane protein (MOMP) gene in C trachomatis while the IDEIA detected genus-specific chlamydial lipopolysaccharide. Discrepant results were resolved by retesting urine specimens with a second (plasmid-based) PCR and taking urethral or endocervical swab results int…

Detection of T-cell receptor delta gene rearrangement by clonal specific polymerase chain reaction.

A sensitive and specific technique to detect minimal residual disease for T-cell malignancies was explored. Southern analysis and polymerase chain reaction (PCR) were used to detect the rearranged V-D-J segment of T-cell receptor delta (TCR delta) gene from malignant cell specimens of patients with leukemia and lymphoma of T-cell lineage. The PCR product was sequenced and from the DNA sequences of the V-D-J region, a 3' antisense primer was designed and synthesized for clonal specific PCR (CS-PCR). Seven of the 22 T-ALL (32%) and 5 of 18 (28%) T-cell lymphoma showed clonal rearrangement by Sou…

Detection of adenovirus DNA in peripheral blood mononuclear cells by polymerase chain reaction assay.

Adenovirus can establish persistent infections which may reactivate and cause disease in immunocompromised hosts. Lymphocytes have been postulated to serve as a site of adenoviral persistence based upon the ability to isolate adenovirus from tonsils and to detect adenovirus DNA by Southern blot hybridization in peripheral blood mononuclear cells (PBMC). To test this hypothesis, a more sensitive and specific polymerase chain reaction (PCR) assay was developed to detect adenovirus DNA. Two sets of nested primers were designed to conserved sequences in the adenovirus E1A and hexon genes. The E1A …

Polymerase chain reaction-based detection of Tropheryma whippelii in central nervous system Whipple's disease.

Whipple's disease of the central nervous system (CNS) may be associated with normal intestinal histology as a result of minimal or patchy in … patients and frozen pontine tissue in 1. Two primer pairs (W3F-W4R, W3F-W2R) were used in separate polymerase chain reactions (PCRs) to amplify fragments of Tropberyma whippelii 16S rDNA from these tissue samples. PCR amplicons were detected only in the duodenal tissues from the 2 patients with OMM. The sequences of these amplicons were identical to the corresponding regi …

Mantle cell lymphoma. Rapid polymerase chain reaction-based genotyping of a morphologically heterogeneous entity.

BACKGROUND: Mantle cell lymphoma is heterogeneous at the morphologic level. Since this B-cell lymphoma may be confused with other entities, ancillary molecular testing may be necessary for definitive diagnosis. A polymerase chain reaction-based method, which is less complicated and more rapid than that generally available for the detection of immunoglobulin heavy chain (IgH) and bcl-1 gene rearrangements, would be helpful in this process. METHODS: Thirty-one mantle cell lymphoma samples (frozen or ethanol-preserved) from 29 patients were studied with two separate polymerase chain reaction ass…

Evaluation of a polymerase chain reaction-based assay for diagnosis of Wuchereria bancrofti infection.

To assess the utility of a polymerase chain reaction (PCR)-based method for diagnosis of Wuchereria bancrofti infection, blood, plasma, and paraffin-embedded tissue samples were tested using a PCR-based assay that detects a W. bancrofti-specific repetitive DNA sequence. The assay was positive in 100 microL of blood from 40 of 42 microfilaria-positive subjects, the 2 subjects with negative assays having microfilarial counts of 1. Samples from 127 uninfected subjects were PCR-negative. The assay was also positive in 7 of 10 daytime samples in regions where infection is nocturnally periodic; PCR …

Rapid detection and typing of circulating mumps virus by reverse transcription/polymerase chain reaction.

The reverse transcription/polymerase chain reaction (RT-PCR) was used to amplify a 129-bp fragment of the mumps virus F gene from strains circulating in the Siena area from 1993-1995. The nucleic acid was amplified directly from the samples; no growth in cell culture was required. Nucleotide sequence analysis and the comparison with other virus strains enabled the typing of the detected viruses. There appears to be more than one lineage of mumps virus circulating at any given time in the same location. A PCR assay coupled with the sequencing of the 5' end of the F gene seems to be a convenient…

[Differentiation of condylomata acuminata from pseudocondyloma by using multiple primer pairs polymerase chain reaction].

Condylomata acuminata were differentiated from pseudocondyloma by using multiple primer pairs polymerase chain reaction (PCR) technique. The paraffin-embedded tissues of 95 cases of condylomata acuminata and 33 cases of pseudocondyloma were detected for HPV DNA. Results show that HPV 6/11 DNA are found in 86/95 (90.5%) cases of condylomata acuminata, and no HPV DNA is detected in 33 cases of pseudocondyloma. The results suggest that pseudocondyloma is not associated with HPV. Multiple primer pairs PCR technique is a preferable method for differentiating condylomata acuminata from pseudocondylo…

Rapid simple and nested polymerase chain reaction for the diagnosis of Pneumocystis carinii pneumonia.

We have developed a rapid and easy extraction procedure for polymerase chain reaction (PCR) protocols. Using this simplified step, we evaluated the sensitivity and the specificity of a simple PCR using the primers of Wakefield et al, and of a nested PCR, using new internal primers selected by us, in a total of 89 bronochoalveolar lavage (BAL) fluid samples from 43 immunosuppressed patients. In 13 patients, Pneumocystis carinii pneumonia (PCP) was diagnosed by immunofluorescent antibody (IFA) staining performed on BAL cells cytospun on microscope slides. In seven of these patients we attempted …

[Studies on tissue distribution and expression of Epstein-Barr virus using polymerase chain reaction].

Epstein-Barr virus (EBV), a widespread human herpesvirus, establishes a life-long carrier state after primary infection. It is known that EBV infects mainly B lymphocytes and oral epithelia in vivo. However, other potential sites of EBV latent and/or permissive infection in human body have not been fully clarified. To investigate this, systemic autopsied tissues from 18 EBV-seropositive individuals without apparent EBV-related diseases were examined for EBV genomic DNA and virus-specific mRNA, by using the polymerase chain reaction technique. EBV DNA was frequently detected in oral mucosa/tong…

Обнаружение серогрупповых вирусов в Калифорнии с использованием универсальных капсюлей и обратной транскрипции-полимеразы.

Были выявлены универсальные капсюли и разработан протокол, пригодный для быстрого обнаружения сложных вирусов калифорнийского энцефалита (СЕ) в цепной реакции обратной транскрипции-полимеразы (RT-PCR). Эти капсюли соответствуют последовательности в областях кодирования G2 glycoprotein среднеразмерного сегмента RNA. Идентификация дополнительных продуктов была подтверждена последовательности в работе клонов или продуктов ПЦР. Этот метод позволяет обнаруживать 40 формирующих табличек (PFU) непосредственно на геле агароза, пропитанном этимидиумом, и чувствительность возрастает до 0,4-1 ПФУ, когда

Совпадение в аннотации оригинала
Universal primers have been identified and a protocol developed that are suitable for rapid detection of California encephalitis (CE) complex viruses in a reverse transcription-polymerase chain reaction (RT-PCR) assay. These primers correspond to sequences in the coding regions of the G2 glycoprotein of the middle-size RNA segment. The identities of the ampl…

[Detection of pathogenic Entamoeba histolytica DNA in liver abscess aspirates with polymerase chain reaction].

Polymerase chain reaction (PCR) was used to diagnose amebic liver abscess by detecting pathogenic Entamoeba histolytica DNA in liver aspirates from patients with liver abscess. Oligonucleotide primers specific for gene encoding of the 30,000 molecule of pathogenic Entamoeba histolytica were used in the test. Liver aspirates from 23 cases with amebic liver abscess as evidenced by typical clinical findings or very high titres of anti-E. Histolytica antibodies with ELISA were found to be positive with PCR. Fourteen control samples (3 cases of bacterial liver abscess, 1 of liver cancer and 10 of a…