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Quantification of interleukin-6 mRNA in wallerian degeneration by competitive reverse transcription polymerase chain reaction.

Following peripheral nerve injury, cytokines might be critically involved in regulating the cellular response within the lesioned nerve but are frequently difficult to measure due to their low abundancy. Competitive quantitative RT-PCR assays are potentially useful in quantifying cytokine mRNAs in small tissue samples but frequently do not fulfill the theoretical demands on proportional amplification of natural and standard sequences. Using this technique, we constructed standard curves for rat interleukin (IL)-6 mRNA with strictly proportional slopes and shifts when varying natural and standa

Enterobacterial intergenic consensus sequence polymerase chain reaction as a typing method for Burkholderia (Pseudomonas) cepacia.

OBJECTIVE: We developed a rapid polymerase chain reaction (PCR) method, which utilizes primers from an enterobacterial repetitive consensus sequence, to differentiate among strains of Burkholderia cepacia. We then applied this method to isolates from a device-associated pseudoinfection and also compared the discriminative typing potential with that of another PCR-based method. METHODS: A simplified lysis procedure was used to generate DNA substrate for amplification by a PCR method which utilized primer ERIC2 and a reverse complement of ERIC1R. The alternative PCR-based method employed primers…

Evaluation of the polymerase chain reaction test for detecting chlamydial DNA in adult chlamydial conjunctivitis.

PURPOSE: A new polymerase chain reaction (PCR) test (Amplicor, Diagnostics, Branchburg, NJ) was evaluated for its ability to detect chlamydial DNA from previously obtained adult conjunctival specimens. METHODS: The sensitivity of this PCR test was determined on 42 adult conjunctival specimens that were culture-positive for Chlamydia. The specificity was determined by testing 40 true-negative specimens that included 10 normal conjunctival samples and 20 ocular specimens that were culture-positive for herpes simplex virus or adenovirus. The remaining ten samples consisted of ocular bacterial pa…

[Direct detection of Toxoplasma gondii with polymerase chain reaction in diagnosis of fetal toxoplasma infection].

Primary infection with Toxoplasma gondii during pregnancy may affect the fetus and result in congenital toxoplasmosis. In Austria serological screening for detection of newly acquired infection during pregnancy was introduced in 1975. In this study we used polymerase chain reaction (PCR) for detection of fetal infection with Toxoplasma gondii. Amniotic fluid samples were analyzed from 11 women with serological indication of acute toxoplasmosis infection. Nine of these women had already received treatment prior to amnio-centesis and no evidence of Toxoplasma gondii DNA was detected with PCR in …

Multiplex polymerase chain reaction for human herpesvirus-6, human cytomegalovirus, and human beta-globin DNA.

Human cytomegalovirus and human herpesvirus-6 are closely related viruses which cause similar diseases, have similar cellular repositories of latent infection, and may be detected largely in the same types of clinical specimens. DNA amplification appears likely to play an increasing role in the diagnosis of recent and remote infection with these agents. A sensitive multiplex polymerase chain reaction was therefore developed for the two viruses and for human beta-globin DNA. Optimization of parameters such as the primers, primer concentrations, magnesium concentration, and buffer constituents w…

Значимость обнаружения энтоновирусной РНК в миокардических тканях посредством обратной транскрипции-полимеразы цепной реакции.

Известно, что у Enterovirus есть связь с человеческим миокардитом и расширенной кардиомиопатией (DCM). Мы проверили образцы биопсии у пациентов с активным миокардитом или DCM, и миокардические ткани из вскрытий без сердечных заболеваний с помощью генной апплификации полимеразы (ПХД). Положительные внтеровиральные сигналы были продемонстрированы 4 из 5 пациентов с активным миокардитом (80%), 7 из 42 пациентов с DCM(17%) и 3 из 27 вскрытий (11%). Из 7 пациентов PCR-положительные с DCM, 3 случая показали гипертрофию и беспорядок кардиоцитов, фиброз в интерститиуме и несколько подстрекательских им

Совпадение в аннотации оригинала
Enteroviruses are known to have association with human myocarditis and dilated cardiomyopathy (DCM). We screened biopsy specimens from patients with active myocarditis or DCM, and myocardial tissues from autopsy cases without cardiac diseases using polymerase chain reaction (PCR) gene amplification. Positive enteroviral signals were demonstrated from 4 of 5 …

Detection of Rickettsia tsutsugamushi specific DNA from the lymphocyte of patients by polymerase chain reaction.

Polymerase chain reaction was used to detect Rickettsia tsutsugamushi specific DNA from the lymphocytes of patients in the acute phase of the disease. The primers used in the PCR were based on the nucleotide sequences of gene encoding of the 56 kDa antigen of the Gilliam strain. The PCR product is a 78 bp fragment which can be hybridized by the 78 bp DNA probe of the Gilliam strain. Comparison of the results obtained by polymerase chain reaction, immunofluorescent assay and cell culture suggests that the polymerase chain reaction method is the most sensitive one among the three in diagnosis of…

Detection of submicroscopic lymph node metastases with polymerase chain reaction in patients with malignant melanoma.

BACKGROUND: The presence or absence of lymph node metastases in patients with malignant melanoma is the most powerful prognostic factor for … METHODS: The assay is accomplished using the combination of reverse transcription and double-round polymerase chain reaction (RT-PCR). The amplified samples are examined on a 2% agarose gel and tyrosinase cDNA is seen as a 207 base pair fragment. Lymph node preparations from 29 patients who were clinically stage I and II and undergoing elective node dissections were analy …

The polymerase chain reaction can reveal the occurrence of naturally mixed infections with Leishmania parasites.

On isolation and characterization of Leishmania parasites from Sudanese patients with visceral leishmaniasis (VL), four cases of mixed infections were found. Three of those cases were from the Eastern Sudan focus of VL. In one case the patient was found to be concomitantly infected with Leishmania donovani and Leishmania aethiopica, while the remaining three patients possessed mixed infections of Leishmania donovani and Leishmania major. Mixed infections were identified by PCR amplification of Leishmania kinetoplast DNA (kDNA) from parasites in culture or in original patient aspirate material

Detection of la-1 oncogene in paraffin-embedded cervical-cancer tissues by polymerase chain-reaction.

The polymerase chain reaction (PCR) technology was used to identify the LA-1 oncogene in cervical cancer cell lines and paraffin embedded tissues. In addition to the products of anticipated size, a 400 bp fragment homologous to the LA-1 oncogene was amplified from 44% of low grade SIL and 60% of invasive cancer tissue. The 400 bp fragment was not amplified from tissues diagnosed as chronic cervicitis, Herpes Simplex Virus (HSV) infection or erythema multiforme. It is suggested that PCR with LA-1 oncogene primers may be a rapid and sensitive diagnostic test for the detection of the SIL patient …

Polymerase chain reaction-based diagnosis of Onchocerca volvulus infection: improved detection of patients with onchocerciasis.

Definitive diagnosis of Onchocerca volvulus (Ov) infection requires the identification of the parasite in either the skin or subcutaneous nodules. These parasitologic approaches suffer from poor sensitivity. To assess the efficacy and utility of a polymerase chain reaction (PCR)-based diagnosis for Ov infection, skin snips were examined from 94 persons in an Ov-endemic region of Ecuador, and results were compared in a blinded fashion with those of a PCR assay based on the Onchocerca-specific repetitive DNA sequence, O-150. All 60 patients microfilaria-positive on skin snip examination were pos…

Cytokine and growth factor expression in Paget's disease: analysis by reverse-transcription/polymerase chain reaction.

We investigated expression of several cytokines and growth factors in explants of Pagetic and non-Pagetic bone samples using the technique of reverse-transcription/polymerase chain reaction (RT/PCR). Transcripts for IL-1 alpha and IL-1 beta, TNF-alpha, TNF-beta, IL-6, basic fibroblast growth factor (bFGF), transforming growth factor beta (TGF-beta) and insulin-like growth factor-I (IGF-I) were found to a variable degree in both Pagetic and non-Pagetic bone samples, but there was no significant difference in the patterns of expression for these factors in Pagetic bone (n = 18) as compared with …

Simple procedure for discrimination and typing of enteric adenoviruses after detection by polymerase chain reaction.

A procedure was developed for specific discrimination and typing of the enteric human adenoviruses, Ad40 and Ad41, after polymerase chain reaction (PCR) amplification of a sequence in the hexon gene highly conserved among all 47 serotypes recognised. By Taq I restriction of the 300 bp amplimers, subgenus F DNA could be discriminated from DNAs of adenoviruses belonging to all other subgenera. Discrimination between Ad40 and Ad41 was subsequently achieved by cleavage with either Cfo I, HinP I, Mae III, Mvn I, and/or Rsa I. Thus, PCR detection of viral DNA combined with restriction analysis of am…

Transmission of varicella-zoster virus from a vaccinee with leukemia, demonstrated by polymerase chain reaction.

A 5-year-old white boy in remission from acute lymphoblastic leukemia who was receiving maintenance anticancer chemotherapy had approximately 200 vesicular skin lesions 1 month after receiving live attenuated varicella vaccine. About 2 to 3 weeks later, a mild illness resembling varicella occurred in his susceptible siblings and in three of his classmates. Vaccine-type varicella-zoster virus was demonstrated by polymerase chain reaction in swab specimens from vesicular lesions in his two siblings, in whom antibody to varicella-zoster virus also developed.

Herpes simplex type 2 in a patient with Mollaret's meningitis: demonstration by polymerase chain reaction.

We describe a patient with recurrent self-limited aseptic meningitis meeting the criteria for Mollaret's syndrome, in whom extensive serological investigation and cerebrospinal fluid analysis failed to reveal a specific cause. Using polymerase chain reaction techniques to amplify herpes simplex viral DNA, we were able to identify the presence of herpes simplex virus type 2 in the cerebrospinal fluid. The duration of clinical illness may have been shortened by treatment with intravenous acyclovir. We suggest that herpes simplex virus may be a more common cause of aseptic meningitis than has bee…

Detection of human papillomavirus by in situ polymerase chain reaction in paraffin-embedded cervical biopsies.

The efficiency of the in situ polymerase chain reaction (PCR) for the detection of human papillomavirus (HPV) DNA sequences in 20 cervical biopsies fixed with buffered formalin, paraffin-embedded and revealed negatively by conventional in situ hybridization (ISH) has been investigated. The biopsies were classified histologically into condylomata acuminata without dysplasia, cervical intraepithelial neoplasia and carcinoma in situ. Amplified HPV DNA was performed after an optimal proteolytic digestion using one pair of consensus oligonucleotide primers located in the L1 ORF of HPV 6 and ISH was…

Detection of Trichomonas vaginalis using the polymerase chain reaction in pregnant and non-pregnant women.

OBJECTIVE: Trichomonas vaginalis vaginal infections are often both asymptomatic and difficult to detect by current methods. We evaluated the ability of a newly developed polymerase chain reaction (PCR) assay to identify T. vaginalis in vaginal samples from pregnant and non-pregnant women. METHODS: In the 1st study, we compared the prevalence of T. vaginalis detection by PCR and culture using Diamond's medium in 52 women with symptoms of vaginal infection. In the 2nd study, T. vaginalis was detected using PCR and wet mount microscopy in 131 asymptomatic pregnant women. RESULTS: Among the wome…

Renin messenger RNA, detected by polymerase chain reaction, can be switched on in rat atrium.

OBJECTIVE: The aim of the present study was to use a highly sensitive, polymerase chain reaction (PCR) technique to examine the controversial question of renin gene expression in the heart and to determine whether expression in cardiac tissue can be stimulated. DESIGN: The study involved female Wistar rats fed either a normal-sodium diet or a low-sodium diet, together with an angiotensin I converting enzyme inhibitor, enalapril, orally for 1 week. METHODS: RNA was extracted from atrium, ventricle, kidney (positive control) and submandibular gland (negative control) and, after reverse transcr…

Identifying human immunodeficiency virus infection at birth: application of polymerase chain reaction to Guthrie cards.

Guthrie cards containing dried blood spots from 67 children now known to be infected with human immunodeficiency virus (HIV) and 63 children now classified as having had seroreversion were retrieved from the newborn infant archives from 1986 through 1991 to determine whether the polymerase chain reaction (PCR) could predict the infection at birth. The PCR assays operating at a sensitivity capable of detecting 2 to 10 copies of HIV proviral DNA per microgram were able to detect HIV proviral DNA in 52% (35/67) of the infected neonatal blood specimens. Longer storage times did not decrease PCR po…

Detection of hepatitis C virus genome in human serum by multi-targeted polymerase chain reaction.

A multi-targeted "hemi-nested" PCR (M-PCR) assay in which the primer pairs derived from the 5' non-coding (5'NC) and the nonstructural protein 3 (NS3) regions of HCV genome were concurrently used for amplification in order to compare the sensitivity and specificity of polymerase chain reaction (PCR) with different primer pairs in detecting hepatitis C virus (HCV) genome. Sera from patients with virus-associated liver diseases were examined for the presence of HCV RNA by the M-PCR method following reverse transcription to cDNA. The amplified products derived from both the 5'NC and the NS3 regio…