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Diagnosis of chronic hepatitis C: comparison of immunoassays and the polymerase chain reaction.

We evaluated a series of first- and second-generation enzyme-linked immunoassays for anti-HCV and compared the findings to those with two confirmatory assays, the recombinant immunoblot assay and serum HCV RNA, in patients with chronic hepatitis C, primary biliary cirrhosis, and chronic hepatitis B. All second-generation immunoassays had good sensitivities (98-100%). Interestingly, detection of HCV RNA had a sensitivity of only 93%, although it was 100% specific. The recombinant immunoblot assay and a peptide-based immunoassay also had good specificity (97% and 100%, respectively), whereas the

[Studies on the detection of Legionella pneumophila by the application of polymerase chain reaction (PCR)].

DNA fragment of Legionella pneumophila, the PCR was performed with a pair of artificial synthesized primer. Results of agarose electrophoresis and EB staining showed that there was a 870 bp band shared by serogroups 1-14 of L.pneumophila. The sensitivity of PCR in detecting Legionella from water was 350cfu/ml, however, the specific DNA probe labeled with 32P was .43cfu/ml by blot hybridization. The positive rate of tissue specimens from infected guinea-pigs with Legionella pneumophila was 83.3% by PCR detection, and only 26.6% by bacteriological culture method. An outbeak caused by Lp10 was ve

Evaluation of the polymerase chain reaction for diagnosis of herpes simplex virus encephalitis.

Cerebrospinal fluid samples from 257 patients with suspected herpes simplex virus encephalitis were prospectively analyzed by herpes simplex virus polymerase chain reaction. The polymerase chain reaction indicated herpes simplex virus encephalitis in 9 serologically proven cases and in 14 additional patients. Increased polymerase chain reaction signals were observed together with more severe neurological symptoms (P < 0.01) and within the first days of acyclovir treatment (P < 0.05).

The polymerase chain reaction for the detection of Borrelia burgdorferi in human body fluids.

OBJECTIVE: To analyze clinical fluids for the presence of Borrelia burgdorferi DNA using the polymerase chain reaction (PCR). METHODS: We utilized a modified, nested PCR to detect the presence of Borrelia DNA in 99 samples of serum, urine, cerebrospinal fluid (CSF), or synovial fluid obtained from 44 patients with various stages of Lyme disease and 47 control subjects. Primer specificity was corroborated by examining 2 DNA data banks, testing against DNA from other organisms, and confirming results with a second set of nested primers. RESULTS: Nested PCR was capable of detecting DNA from few…

Comparison of polymerase chain reaction with culture and enzyme immunoassay for diagnosis of pertussis.

A polymerase chain reaction (PCR) assay amplifying a segment of a repeated gene element of Bordetella pertussis was compared with culture and enzyme immunoassay (EIA) for the diagnosis of pertussis. The PCR assay was specific for B. pertussis in tests with a panel of other bacteria and with an extensive collection of specimen material from healthy persons and children with respiratory infections other than pertussis. The PCR assay was used in the analysis of 117 nasopharyngeal swabs collected from children at an elementary school at which a pertussis outbreak occurred. Fifty-six (48%) of the 1…

Utility of the polymerase chain reaction (PCR) for prenatal diagnosis of genetic disease.

Gene amplification by the polymerase chain reaction (PCR) has been applied to prenatal diagnosis for alpha and beta thalassemias (1 and 5 cases respectively), Hemoglobin (Hb) Lepore/beta thalassemia (1 case) and cystic fibrosis (14 cases). Chorionic villus samples were obtained in the tenth week of pregnancy and DNA analysed in parallel with conventional gene mapping. Direct diagnosis of the common Mediterranean beta-thalassemia mutations (IVS-1-110 and codon 39), Hb Lepore, and the delta F508 mutation causing cystic fibrosis was achieved by hybridization of amplified material with pairs of al…

Detection of Mycoplasma genitalium in the genitourinary tract of women by the polymerase chain reaction.

A polymerase chain reaction (PCR) was used to demonstrate the presence or absence of Mycoplasma genitalium in the lower genital tract of 57 women who attended a sexually transmitted diseases clinic. The mycoplasma was detected in the cervix of 10 (17.5%) women and also in the vagina of 4 (16%) and the urethra of 6 (24%) of 25 women from whom multiple samples were obtained. Chlamydia trachomatis was detected also by a PCR in 9 (16%) of the women, but only 3 were chlamydia-positive and mycoplasma-positive. M. genitalium was detected occasionally in women with vaginal disease (for example, bacter…

DNA sequences for the specific detection of Cryptosporidium parvum by the polymerase chain reaction.

The objective of this project was to construct specific and sensitive molecular probes and amplification primers for Cryptosporidium parvum that could be used in diagnosis, retrospective tissue studies, and in epidemiologic surveys. Whole genomic DNA was extracted from oocysts of C. parvum purified from human and bovine feces. A genomic library was constructed in plasmid pUC18 and propagated in Escherichia coli DH5 alpha. Transformants were screened by colony hybridization and autoradiography. The 2.3-kilobase segment in plasmid pHC1, a clone specific for C. parvum, was sequenced by the Sanger

Localization of breakpoints by polymerase chain reactions in Burkitt's lymphoma with 8;14 translocations.

Translocations involving chromosomes 8 and 14 in Burkitt's lymphoma (BL) often involve the switch mu (Smu) region on chromosome 14, which contains multiple repeats. This has enabled us to use the polymerase chain reaction (PCR) to detect breakpoints that involve this region on chromosome 14 and the c-myc gene on chromosome 8. Using pairs of flanking primers, each pair including one annealing to repeat sequences within the switch region and one of three primers from the c-myc region (first intron, 3', or 5' flanking sequence of the first exon of c-myc), we have been able to amplify DNA fragment…

Identification of Toxoplasma gondii in paraffin-embedded sections by the polymerase chain reaction.

We used the polymerase chain reaction to amplify DNA fragments specific to Toxoplasma gondii. The sensitivity of the technique allowed for the detection of as few as ten cultured T. gondii tachyzoites. We applied the same amplification technique to deparaffinized ocular sections from two cases of ocular toxoplasmosis. Although toxoplasmic cysts could only be seen in one eye by optical microscopy, polymerase chain reaction allowed the identification of the parasite in both cases. Our study indicates the feasibility of a sensitive DNA-based assay to complement pathologic studies of an ocular par…

Обнаружение STAT4 у пациентов с множественным склерозом с помощью цепной реакции полимеразы и потокцитометрии.

Множественный склероз (MS) — болезнь центральной нервной системы, характеризующаяся постепенным обезвоживанием и воспалением. MS характеризуется атаками иммунной системы на миелиновую оболочку вокруг нервных волокон. Исследования ассоциации геномов выявили полиморфизм в преобразователе сигнала и активаторе транскрипции 4 (STAT4) ген, повышающий риск для MS. Этот полиморфизм влияет на клетки Т-аппер1 (Th1) для отделения интерферона-гаммы цитокина, стимулируемого интерлейкином (IL)-12. Цель этого исследования состояла в том, чтобы определить связь активного заболевания MS с генотипами STAT4, обн

Совпадение в аннотации оригинала
Multiple sclerosis (MS) is a disease of the central nervous system, characterized by progressive demyelination and inflammation. MS is chara … study aimed to determine the association of MS active disease with STAT4 genotypes, detected by the polymerase chain reaction (PCR) and STAT4 protein level detected by flowcytometry. The study included 80 MS patients, and 70 controls matched for age and gender. We used the restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR) and flowcytometry to de …

Polymerase chain reaction in the diagnosis of invasive aspergillosis: approaches for appropriate use.

Detection of aspergillus by PCR is a helpful tool for early diagnosis. The test has excellent sensitivity and specificity with a high negative predictive value. Well-accepted, standardized method for DNA extraction for PCR testing is to be adopted for all commercial assays and conclusive validation data are awaited in varied clinical settings. This perspective offers guidance for utilizing PCR testing while awaiting such data. Quantification by PCR, species-specific identification assays and detection of resistance genetic markers are of future promise. Herein, we summarize the available data

The Utility of Nonroutine Intraocular Fluid Polymerase Chain Reaction for Uveitis in Indonesia.

PURPOSE: To investigate the utility of nonroutine polymerase chain reaction analysis of intraocular fluid to guide the diagnosis of infectious uveitis. PATIENTS AND METHODS: A retrospective cohort study was conducted by reviewing medical record data from intraocular fluid samples of uveitis patients who underwent single-plex real-time polymerase chain reaction analysis at the Department of Ophthalmology, Faculty of Medicine, Universitas Indonesia - Cipto Mangunkusumo Kirana Eye Hospital between January 2014 and December 2018. RESULTS: The positivity rate of nonroutine polymerase chain reacti…

Multiplex bacterial polymerase chain reaction in a cohort of patients with pleural effusion.

BACKGROUND: The identification of the pathogens in pleural effusion has mainly relied on conventional bacterial culture or single species polymerase chain reaction (PCR), both with relatively low sensitivity. We investigated the efficacy of a commercially available multiplex bacterial PCR assay developed for pneumonia to identify the pathogens involved in pleural infection, particularly empyema. METHODS: A prospective, monocentric, observational study including 194 patients with pleural effusion. Patients were evaluated based on imaging, laboratory values, pleura ultrasound and results of tho…

Cutaneous Leishmaniasis of Lip and Role of Polymerase Chain Reaction: A Case Report.

The diagnosis of cutaneous leishmaniasis is mostly confirmed by the identification of parasite in a skin smear or biopsy. However, this method may not always be sensitive enough to detect the disease when parasitic load is low. Molecular test such as polymerase chain reactions can be useful in such circumstances. Here, we report a case of cutaneous leishmaniasis diagnosed by a polymerase chain reaction test when both smear and biopsy failed to confirm the diagnosis. A 17-years-old female from mountainous district of Nepal, presented with a crusted plaque over the upper lip for a duration of 6 …

Cytomegalovirus Identification in Blood and Urine of Newborns by Nested Polymerase Chain Reaction.

AIM: To study the frequency of congenital cytomegalovirus (CMV) infection in newborns admitted to the Division of Neonatology, using nested polymerase chain reaction (PCR) and DNA to detect differences in blood and urine specimens. METHODS: The study was carried out for eight months. Newborns (n = 520) hospitalized in five hospitals in Campo Grande, Mato Grosso do Sul, Brazil, were checked for CMV by analysing blood and urine samples. RESULTS: Cytomegalovirus was PCR positive in 13 urine and 10 blood samples. Of the 13 positive urine patients, three (23%) had no clinical signs suggestive of …

Survey of pathogenic Leptospira in rats by polymerase chain reaction in Sisaket Province.

BACKGROUND: Leptospirosis, a zoonotic disease caused by Leptospira, has been a health problem in Thailand for several years. It can be trans … irosis in Sisaket Province. MATERIAL AND METHOD: DNA samples were isolated from rats' kidneys. The polymerase chain reaction (PCR) technique was used for the detection of 16s rRNA and LipL32 genes specific to genus and pathogenic Leptospira, respectively. RESULTS: In areas where patients with leptospirosis had been reported, 8.7% (4/46) of rats were infected with pathoge …

Design, optimization and evaluation of a polymerase chain reaction for detection of borrelia spp.

BACKGROUND: Lyme borreliosis and relapsing fever are important zoonotic diseases worldwide and the improvement of diagnostic strategies is a prioritized task considering the morbidity of these diseases in some areas. PCR based methods appear to be of utmost importance because of the high sensitivity and specificity of these assays. OBJECTIVES: To obtain a molecular method based on PCR for the detection of the genus Borrelia infection in different specimens. RESULTS: Sets of reported primers were evaluated "in silico" and they did not fulfill the proposal parameters. On the other hand, the tw

Increased detection of cutaneous leishmaniasis in Norway by use of polymerase chain reaction.

Cutaneous leishmaniasis (CL) is a parasitic infection and occurs in tropical and subtropical regions worldwide and in the region of the Mediterranean Sea. The diagnosis is based on the clinical appearance and biopsy findings that may be supplemented with polymerase chain reaction (PCR). In this study 20 cases were selected if (i) a histopathological diagnosis of granulomatous dermatitis was made, (ii) CL was taken into consideration or (iii) the diagnosis was CL. PCR analysis with primers specific for leishmania was performed on archived histological specimens and was positive in 6 of the 20 c…

Role of reverse transcriptase polymerase chain reaction for the diagnosis of human rabies.

BACKGROUND & OBJECTIVES: Traditionally, rabies diagnosis is made by demonstration of rabies viral antigen by direct immunofluorescence (DIF) and mouse inoculation test (MIT). The present study was carried out to evaluate the role of reverse transcriptase-polymerase chain reaction (RT-PCR) in comparison with these conventional techniques for the diagnosis of rabies. METHODS: Skin biopsies, corneal impression smears and saliva sample were collected ante-mortem and brain tissue and CSF were collected post-mortem from ten clinically suspected rabies patients. DIF, Seller staining, MIT and RT-PCR …