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Identification of the forest strain of Onchocerca volvulus using the polymerase chain reaction technique.

Annual mass treatment with ivermectin for 12-15 years in endemic communities is the control strategy adopted by the African Programme for Onchocerciasis Control (APOC) for the control of onchocerciasis in Nigeria. This long-term treatment necessitates the use of Polymerase Chain Reaction (PCR) for the proper identification of the Onchocerca species and strains in endemic areas and also for monitoring recrudescence of infection in areas where infection has been controlled. This study, which forms part of a larger study on transmission of onchocerciasis identifies the Onchocerca volvulus strain …

Real time polymerase chain reaction: a new powerful tool for the diagnosis of neurobrucellosis.

BACKGROUND/METHODS: We compared the diagnostic yield of a real time polymerase chain reaction (PCR) assay in cerebrospinal fluid (CSF) samples with conventional microbiological techniques for the diagnosis of neurobrucellosis. Following amplification of a 223 bp sequence specific for Brucella genus, melting curve analysis was performed to verify the specificity of the PCR products. RESULTS: All six patients with neurobrucellosis (three meningitis and three meningoencephalitis) had a positive real time PCR assay, whereas CSF cultures and Wright seroagglutination tests were positive in only two…

Detection of Brucella DNA in sera from patients with brucellosis by polymerase chain reaction.

To overcome the limitations of current diagnostic methods for brucellosis, accurate and simple methods must be sought. In this study, sera of patients diagnosed with brucellosis were examined by polymerase chain reaction (PCR) for the detection of Brucella DNA. Results showed that an amplicon of 223 bp was obtained in 96% (24/25) of tested sera using primers derived from the nucleotide sequence of a gene encoding the 31-kDa Brucella abortus antigen. However, when serum DNA templates were examined by another PCR, an amplicon of 731 bp was obtained in 60% (15/25) of the tested samples using Bruc…

[Assessment of polymerase chain reaction (PCR) to diagnose brucellosis in a Brucella infected herd].

The diagnosis of bovine brucellosis using PCR in blood and milk samples from two dairy herds were compared to in vitro isolation, complement fixation test (CF), competitive ELISA (C-ELISA) in serum, and indirect ELISA (I-ELISA) in milk. Samples were obtained from 99 cows vaccinated with Brucella abortus strain 19, from a naturally infected herd (A), whose cows were also vaccinated with B. abortus strain RB51 as adults, and 100 from brucellosis free herd (B). In herd A, PCR identified 14 B. abortus infected cows: nine infected with wild type, and five with wild type and RB51, B. abortus S 19 wa

Development of a quantitative real-time polymerase chain reaction assay specific for Orientia tsutsugamushi.

Two specific and sensitive polymerase chain reaction (PCR) assays were developed to detect and quantitate Orientia tsutsugamushi, the agent of scrub typhus, using a portion of the 47-kD outer membrane protein antigen/ high temperature requirement A gene as the target. A selected 47-kD protein gene primer pair amplified a 118-basepair fragment from all 26 strains of O. tsutsugamushi evaluated, but it did not produce amplicons when 17 Rickettsia and 18 less-related bacterial nucleic acid extracts were tested. Similar agent specificity for the real-time PCR assay, which used the same primers and …

Whipple's disease: the first Japanese case diagnosed by electron microscopy and polymerase chain reaction.

A 52-year-old man presented with diarrhea and 20 kg weight loss in one year. Enteroscopy showed diffuse yellow-white shaggy mucosa in the duodenum and jejunum. Biopsies of the duodenal mucosa revealed massive infiltration within the lamina propria by foamy macrophages strongly positive for periodic acid-Schiff stain. Electron microscopy demonstrated numerous bacilli within macrophages of the lamina propria. Tropheryma whipplei DNA was detected by polymerase chain reaction. The definitive diagnosis of Whipple's disease was made. Antibiotic therapy dramatically improved his clinical picture. Thi…

Prognostic significance of reverse transcriptase-polymerase chain reaction-negative sentinel nodes in malignant melanoma.

BACKGROUND: Polymerase chain reaction (PCR) is a molecular biology technique that can detect a single metastatic cell in 10(6) to 10(7) normal cells. Its use has been proposed as an additional new method for the detection of malignant melanoma nodal metastases in the sentinel lymph node (SLN) to improve the detection rate guaranteed so far by standard histology (hematoxylin and eosin; H&E) and immunohistochemistry (IHC). METHODS: Since October 1995, 137 patients with primary cutaneous melanoma (Breslow thickness,.75-4 mm) have undergone surgery for selective lymphadenectomy. To identify the S…

Molecular detection of Babesia equi in infected and carrier horses by polymerase chain reaction.

Twenty-three blood samples were used in this study; five were from five naturally infected horses with Babesia equi (B. equi), while eighteen were from asymptomatic horses with equine babesiasis from different localities in Egypt. All samples were subjected to microscopic examination, indirect fluorescent antibody test (IFA) and polymerase chain reaction (PCR). The carrier animals were microscopically detected in 7 out of 18 samples (38.8%) and in 9 of 18 by using IFA (50%), whereas PCR revealed that 14 samples were positive (78%). Two synthetic oligonucleotide primers, based on the B. equi me…

Rapid polymerase chain reaction-based confirmation of cat scratch disease and Bartonella henselae infection.

CONTEXT: Cat scratch disease (CSD) commonly occurs secondary to Bartonella henselae infection, and the diagnosis has traditionally been made by microscopic findings, the identification of organisms by cytochemistry, and clinical history. However, cytochemical analysis tends to be very difficult to interpret, and histology alone may be insufficient to establish a definitive diagnosis of CSD. OBJECTIVE: To demonstrate the presence of B henselae in tissue suspected of involvement by CSD, using a novel polymerase chain reaction (PCR) assay. DESIGN: Isolates of B henselae (American Tissue Culture…

Perinatal screening for group B streptococci: cost-benefit analysis of rapid polymerase chain reaction.

OBJECTIVE: To evaluate the costs and benefits of a group B streptococci screening strategy using a new, rapid polymerase chain reaction test in a hypothetical cohort of expectant mothers in the United States. DESIGN: Cost-benefit analysis using the human capital method. We developed a decision model to analyze the costs and benefits of a hypothetical group B streptococci screening strategy using a new, rapid polymerase chain reaction test as compared with the currently recommended group B streptococci screening guidelines-prenatal culture performed at 35 to 37 weeks or risk-factor-based strat…

Determination of alpha-1 antitrypsin genetic deficiency in duodenal ulcer by polymerase chain reaction.

BACKGROUND: Smoking is one of the most important public health problems among young people. Potential risk factors that may cause vulnerability to smoke in youth should be well known and investigated. The aim of the present study was to evaluate the associations of current smoking behavior and future smoking intention with high-risk personality traits for substance abuse in a clinical sample of Turkish adolescents, and also evaluate nicotine dependence and smoking characteristics with the personality traits in a subsample of regular smokers. METHODS: A cross-sectional study was adopted in whi

Detection of herpesvirus genomes by polymerase chain reaction in cerebrospinal fluid and clinical findings.

BACKGROUND: The viruses of the Herpesviridae family, in particular herpes simplex virus types 1 (HSV-1) and 2 (HSV-2), cytomegalovirus (CMV) … appropriately, as treatment is specific for the particular virus concerned. OBJECTIVES: To apply a polymerase chain reaction (PCR) technique for the diagnosis in a single reaction of the six herpesviruses most frequently detected in the cerebrospinal fluid (CSF) and to analyse clinical events in patients presenting positive results in PCR for herpesviruses. STUDY DESIGN: …

Development of a polymerase chain reaction method for diagnosing Babesia gibsoni infection in dogs.

A pair of oligonucleotide primers were designed according to the nucleotide sequence of the P18 gene of Babesia gibsoni (B. gibsoni), NRCPD strain, and were used to detect parasite DNA from blood samples of B. gibsoni-infected dogs by polymerase chain reaction (PCR). PCR was specific for B. gibsoni since no amplification was detected with DNA from B. Canis or normal dog leucocytes. PCR was sensitive enough to detect parasite DNA from 2.5 microl of blood samples with a parasitemia of 0.000002%. PCR detected parasite DNA from 2 to 222 days post-infection in sequential blood samples derived from …

Rapid and sensitive diagnosis of human adenovirus infections by a generic polymerase chain reaction.

A new adenovirus specific nested polymerase chain reaction (PCR) method is described. It was designed inside the hexon protein gene of the adenovirus genome, and was able to detect DNA of all 47 human adenovirus types in a wide range of clinical samples. A sensitive internal control system able to assure proper analytical conditions for the amplification of as few as 100 molecules of a heterologous DNA was included to avoid false negative results. Sensitivity was estimated at about 10 molecules per tube of a plasmid containing an insert of the first amplification product. The method was able t…

Mosquito surveillance and polymerase chain reaction detection of West Nile virus, New York State.

West Nile (WN) virus was detected in the metropolitan New York City (NYC) area during the summer and fall of 1999. Sixty-two human cases, 7 fatal, were documented. The New York State Department of Health initiated a departmental effort to implement a statewide mosquito and virus surveillance system. During the 2000 arbovirus surveillance season, we collected 317,676 mosquitoes, submitted 9,952 pools for virus testing, and detected 363 WN virus-positive pools by polymerase chain reaction (PCR). Eight species of mosquitoes were found infected. Our mosquito surveillance system complemented other …

Обнаружение арбовирусной РНК непосредственно от москитных однородностей путем обратной транскрипции - полимеразы.

Многие трансмиссивые артроподами (арбовирусы) являются важными патогенами человека с медицинской точки зрения. Разработка эффективного метода обнаружения вирусов с помощью амплификации нуклеиновой кислоты, как, например, цепная реакция полимеразы (ПХД), способствует не только клиническому диагностике, но и вирологическому наблюдению за переносчиками комаров на местах. В этом исследовании описана разработка усовершенствованного и упрощенного анализа для выявления комаров, инфицированных вирусом энцефалита восточной экипиды (EEE), вирусом Cache Valley (CV) и серогрупповыми вирусами Калифорнии (C

Совпадение в аннотации оригинала
Many arthropod-borne viruses (arboviruses) are important human pathogens medically. The development of an effective technique to detect the viruses by using nucleic acid amplification, such as polymerase chain reaction (PCR), improves not only clinical diagnosis but also virologic surveillance of mosquito vectors in the field. In this study, the development …

Enzyme immunoassay for the diagnosis of cat-scratch disease defined by polymerase chain reaction.

Whole-cell immunofluorescent antibody (IFA) tests for detection of anti-Bartonella henselae immunoglobulin (Ig) G are commonly used to diagn … 84 patients with definite CSD (regional lymphadenitis, cat contact, and > or =1 confirmatory test: polymerase chain reaction, skin test, or B. henselae culture). Although this method has been used as a diagnostic tool in several case reports, it has not previously been evaluated in a large study of definitively proven CSD cases. Results of this study indicate that the EIA …

Improved detection of microorganisms by polymerase chain reaction in delayed endophthalmitis after cataract surgery.

OBJECTIVE: To evaluate whether the use of polymerase chain reaction (PCR) improves the identification of the causative pathogen in eyes developing delayed endophthalmitis after cataract surgery. DESIGN: Prospective, noncomparative case series. PARTICIPANTS: Consecutive series of 25 eyes with the clinical diagnosis of delayed endophthalmitis after cataract. MAIN OUTCOME MEASURE: Presence of bacterial or fungal DNA in aqueous humor and vitreous samples. RESULTS: In the aqueous humor the causative pathogen was identified in 84% (n = 21) of the eyes by PCR compared with 0% by diagnostic cultur…

Polymerase chain reaction for detection of Mycobacterium tuberculosis in epiretinal membrane in Eales' disease.

PURPOSE: Tuberculous etiology has been suggested in Eales' disease. Because epiretinal membrane (ERM) is formed on the inner surface of the retina in Eales' disease, it could be the most appropriate intraocular specimen for investigation. Therefore, a nested polymerase chain reaction (nPCR), which detects MPB64 gene of Mycobacterium tuberculosis on the archival specimens of ERM of well-documented Eales' and non-Eales' patients, was applied and the results compared. METHODS: nPCR technique was standardized, and the sensitivity and specificity of the primers were determined. nPCR technique was …