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Development and In-House Validation of a Real-Time Polymerase Chain Reaction for the Detection of Listeria monocytogenes in Meat.

Listeriosis is a foodborne disease caused by Listeria monocytogenes. The aims of this work were to develop and validate an in-house real-time polymerase chain reaction (RT-PCR) for the detection of L. monocytogenes, and to determine its prevalence in raw ground beef samples from 53 butcheries that also sell ready-to-eat foods. One set of primers and one hydrolysis probe were designed for hly gene detection and then challenged with pure strains. The detection was successful for all L. monocytogenes strains analyzed and negative for all non-L. monocytogenes strains (detection limit, 10 colony fo…

Use of the polymerase chain reaction for the diagnosis of asymptomatic Leishmania infection in a visceral leishmaniasis-endemic area.

The diagnosis of asymptomatic infection with Leishmania (Leishmania) chagasi has become more important over recent years. Expansion of visce … in a visceral leishmaniasis endemic area were evaluated clinically and submitted to genus-specific polymerase chain reaction (PCR), serological testing, and the Montenegro skin test. Forty-nine (32.9%) of the subjects had a positive PCR result and none of them developed the disease within a follow-up period of three years. No association was observed between the results o …

Comparative study of microscopy and polymerase chain reaction for the diagnosis of suspected visceral leishmaniasis patients in Nepal.

BACKGROUND: Visceral leishmaniasis is potentially fatal protozoan diseases caused by Leishmania donovani. Nepal is an endemic region in which visceral leishmaniasis causes a major public health problem in the lowland areas that border the endemic areas of Bihar state in India. Accurate diagnosis to inform treatment is a first step in achieving the goal of visceral leishmaniasis elimination from South East Asian regions by 2020. OBJECTIVE: The objective of the present study was to compare between the Microcopy and polymerase chain reaction for diagnosis of visceral leishmaniasis. METHODS: In …

Diagnosis of intraocular lymphosarcoma in a dog by use of a polymerase chain reaction assay for antigen receptor rearrangement.

CASE DESCRIPTION: A 12-year-old castrated male mixed-breed dog was evaluated because of blepharospasm and blindness affecting both eyes. CLINICAL FINDINGS: During examination and diagnostic testing of the dog, fine-needle aspirates of splenic nodules were examined microscopically and stage Vb multicentric large-cell lymphosarcoma was identified. Aqueocentesis was performed, and sample analysis revealed intraocular lymphosarcoma; B-cell neoplasia was confirmed by use of a PCR assay for antigen receptor rearrangement (PARR) performed on samples of aqueous humor. Secondary uveitis and glaucoma w

[The development of method of detection of human pathogenic orthopoxviruses on the basis of polymerase chain reaction in real time].

The set of specific oligonucleotide primers and hybridization testers with the support of TaqMan in real time is proposed to identify and differentiate such human pathogenic orthopoxviruses as smallpox virus, monkey smallpox virus, cowpox virus, variolovaccine. The analytical specificity of the set is determined upon the example of 20 strains of 6 types of human pathogenic orthopoxviruses and made up 100%. The sensibility is established using recombinant plasmids containing orthopoxviruses sequence. The minimal revealed amount of plasmid made up to 20 copies for smallpox and monkey smallpox vi

Evaluation of polymerase chain reaction as an additional tool for the diagnosis of low-intensity Schistosoma mansoni infection.

The aim of the present study was to evaluate polymerase chain reaction (PCR) as an alternative tool for diagnosing schistosomiasis in individuals with low-level parasite burden from areas of low endemicity or under occasional risk of infection by Schistosoma mansoni. A total of 102 samples were tested in this study using 2 PCR assays utilizing distinct primer pairs. One of the primer pairs was targeted to a highly repeated 121-base pair sequence of S. mansoni, and the other was targeted to Schistosoma 28S rDNA. The samples were divided into 4 groups according to parasite burden of the individu…

Development of a multiplex polymerase chain reaction for detection and typing of major human herpesviruses in cerebrospinal fluid.

Infections of the central nervous system (CNS) represent a difficult diagnostic problem for both clinicians and microbiologists. In particul … nal methods for detection of herpesviruses in CNS infections exhibit several drawbacks, whereas the polymerase chain reaction (PCR) on cerebrospinal fluid has revolutionized the neurovirology and is becoming an essential part of the diagnostic work-up of patients with suspected CNS viral infections. A sensitive multiplex PCR method was developed for the simultaneous detect …

Identification of pathogenic Leptospira strains in tissues of a premature foal by use of polymerase chain reaction analysis.

Studies were carried out to determine the cause of death in a prematurely born Thoroughbred foal that died 24 hours after birth. Necropsy revealed gross lesions suggestive of septicemia. A commercial Leptospira polymerase chain reaction (PCR) assay designed to specifically amplify the hemolysis-associated protein 1 (hap1) gene present only in pathogenic Leptospira strains detected the presence of Leptospira DNA in various tissues of the foal. Histologic examination of lung, liver, kidney, and myocardium revealed numerous spirochetes in Warthin-Starry-stained tissue sections. Results of PCR ana…

Resolving T-cell receptor clonality in two and genotype in four multiplex polymerase chain reactions.

BACKGROUND AND OBJECTIVES: The diagnosis of T-cell neoplasia requires the use of immunohistochemistry on tumor sections or molecular genetic analysis of T-cell receptor (TCR) clonality. Multiplex polymerase chain reactions (PCR) offer a sensitive and expeditious approach to determining clonality early in the diagnostic work-up. We determined the sensitivity and specificity of four multiple PCR for genotyping lymphoid neoplasms at the TCR loci gamma (TCRG), delta (TCRD) and beta (TCRB, including complete [Vbeta-Jbeta] and incomplete [Dbeta-Jbeta] rearrangements). DESIGN AND METHODS: Template D…

Molecular assay for the detection of Cochlosoma anatis in house flies and turkey specimens by polymerase chain reaction.

A 1520 bp region of Cochlosoma anatis mtDNA 16S gene was subjected to DNA sequencing and a 466 bp portion was compared with other protozoan 16S sequences to develop PCR primers specific for C. anatis. This PCR diagnostic method allowed identification of C. anatis from house flies, Musca domestica L., turkey gut, and fecal samples within 6 h after field-collected samples reached the laboratory. House flies detected carrying C. anatis using the diagnostic 374 bp amplicons represented the first record of this protozoan in house flies.

Antitumor effect of radioimmunotherapy in a mouse model of testicular tumor with micrometastases defined by polymerase chain reaction.

The efficacy of radioimmunotherapy (RIT) was evaluated by using a mouse model of testicular tumor with macro- and micrometastases to various organs. RIT consisting of a single intravenous injection of 5.8 MBq of I-131-labeled anti-placental alkaline phosphatase (PLAP) MAb was conducted one day or 7 days after testicular implantation in SCID mice of HeLa Hep2 cells expressing PLAP. RIT antitumor effect was significant for primary tumors as well as micrometastases defined by polymerase chain reaction (PCR) assay. However, ablation of tumor cells could be achieved with this treatment only at the …

Isolation and amplification by polymerase chain reaction DNA of Babesia microti and Babesia divergens in ticks in Poland.

Babesia microti and B. divergens, the etiological agents of human babesiosis, are transmitted by the bite of Ixodes ricinus. The purpose of this study was differentiation of those two species in ticks collected in urban woods in the city Szczecin (north-western Poland). The prevalence of the DNA of Babesia were investigated by PCR amplification with primers to the fragment from a gene encoding the nuclear small-subunit ribosomal RNA (SS-rDNA). We examined a total of 533 specimens of Ixodes ricinus. The mean infection rate was 16.3%. Our results indicate that a B. microti and B. divergens--spec

[A pilot study on the value of duplex polymerase chain reaction method in early diagnosis of Legionella pneumonia].

OBJECTIVE: To investigate the value of duplex polymerase chain reaction (DPCR) in early diagnosis of Legionella pneumonia by detecting Legionella DNA in sputum and bronchoalveolar lavage fluid(BALF). METHODS: During the process of DPCR, two different sets of oligonucleotide primers were simultaneously used to amplify 386bp 16SrRNA gene fragment and 206bp mip gene fragment. These two primers were designed according to the sequences of 16SrRNA gene and mip gene of Legionella. The sputum and BALF of patients from two groups, including a Legionella pneumonia group (n = 15) and an ordinary pneumon…

The detection of viral genomes by polymerase chain reaction in the myocardium of pediatric patients with advanced HIV disease.

OBJECTIVES: The aim of this study was to investigate the frequency of viral nucleic acid detection in the myocardium of human immunodeficien … ection. The hearts were examined histologically and analyzed for the presence of viral sequences by polymerase chain reaction (PCR) or reverse transcription-PCR. RESULTS: Myocarditis was detected histologically in 11 of the 32 HIV-infected patients, and borderline myocarditis was diagnosed in another 13 cases. Infiltrates were confined to the epicardium in two additional …

[Evaluation of the polymerase chain reaction technique for the diagnosis of meningitis caused by Neisseria meningitidis and Haemophilus influenzae].

BACKGROUND: Bacterial meningitis is a severe infection of the central nervous system (CNS), most frequently caused by Neisseria meningitidis in our setting. Microbiologic diagnosis of bacterial meningitis is not enough sensitive because its efficiency can be affected by the therapeutic regimen given to the patient. Polymerase chain reaction (PCR) can provide a more sensitive diagnosis and allow us to get an earlier result. OBJECTIVES: To assess the sensitivity and specificity of a PCR technique for the diagnosis of meningitis caused by N. meningitidis and Haemophilus influenzae. MATERIAL AND…

[The detection of cytomegalovirus DNA by the polymerase chain reaction in different biological materials in patients with organ allografts].

Ninety-eight patients with organ allografts were examined using the polymerase chain reaction (PCR). Cytomegalovirus (CMV) DNA was not found in the blood of any of 67 blood donors. At least 85% patients with CMV DNA in the plasma developed CMV disease. CMV DNA was detected in leukocytes of 22 (40.7%) out of 54 patients with negative results of analysis for CMV DNA in the plasma; 16 (29.6%) of these 22 presented with clinical symptoms of CMV infection by the moment of investigation. Results of PCR analysis of blood, urine, and saliva specimens of 30 patients examined did not always coincide. DN…

[Detection of Toxoplasma gondii DNA in the autopsy cases of abnormal birth and teratosis by polymerase chain reaction].

Toxoplasma gondii DNA in the autopsy tissues of 72 cases of the premature, stillbirth and congenital malformation was detected using polymerase chain reaction (PCR). The specific amplified products were found in 9 cases, i.e. one case of premature, 3 cases of stillbirth, one case of hydrocephalus and 4 cases of other teratosis. The positive rate was 12.5%. Ten control samples were negative. The results showed that Toxoplasma gondii infection was present in the disease cases. The finding indicates that the infection of Toxoplasma gondii may be one of the main causes of the abnormal birth and th…

Тройной вирусный ретинит, диагностированный полимеразной цепной реакцией витроусовой биопсии у пациента с синдромом Рихтера.

ЦЕЛЬ: Сообщить об оценке и идентификации вирусов герпеса, связанных с ретинитом у пациента с синдромом Рихтера. МЕТОДЫ: Диагностическая витрэктомия была проведена на пациенте с системной лейкемией и ретинитом. Витриновая проба оценивалась цитологией, анализом цитокина ЭЛИСА и обнаружением вируса при цепной реакции полимеразы. Результаты: Витриновый биопсический образец не показал злокачественных клеток, но преобладал CD8+ лимфоцит, инфильтрированный с повышенным интерфероном гамма- и интерлейкин-6. Анализ ДНК и южного пятна показал ДНК герпеса simpex, варицелла-зостера и цитомегаловируса.ЗАКЛЮ

Совпадение в аннотации оригинала
PURPOSE: To report the evaluation and identification of herpes viruses associated with retinitis in a patient with Richter syndrome. METHODS: Diagnostic vitrectomy was performed on a patient with systemic leukemia and retinitis. The vitreous sample was evaluated by cytology, analysis of cytokines by ELISA, and detection of virus by polymerase chain reaction…

[Detection of mycobacterial DNA with polymerase chain reaction in eye discharge and gastric juices in a case of scleritis].

We report a case of mycobacterial scleritis in which prompt diagnosis was made by the detection of mycobacterial DNA with polymerase chain reaction (PCR) in eye discharge and gastric juices, when conventional tests were negative. A 77-year-old woman who had a past history of pulmonary tuberculosis visited the outpatient clinic of Tokai University Hospital complaining of pain in her right eye. She was diagnosed as having scleritis and uveitis. There were no indications of active tuberculosis. We examined the gastric juices, sputum, and eye discharge by microscopy, culture, and PCR for detection…