scholarly journals Feline immunodeficiency virusinfection: a comparative study of different diagnostic techniques

2004 ◽  
Vol 56 (1) ◽  
pp. 13-18
Author(s):  
E. Mortola ◽  
G. Oliva ◽  
M. Risso ◽  
M. Pecoraro ◽  
M.C. Venturini
Keyword(s):  
High Sensitivity ◽  
Epidemiological Survey ◽  
Epidemiological Studies ◽  
Diagnostic Techniques ◽  
Immunofluorescence Assay ◽  
Feline Immunodeficiency Virus Infection ◽  
Immunodeficiency Virus ◽  
Pcr Method ◽  
Specific Reactivity ◽  
Polymerase Chain

This study evaluated an indirect immunofluorescence assay (IFA) to detect feline immunodeficiency virus infection (FIV) antibody in a comprehensive epidemiological survey of FIV in Argentina. IFA modified in our laboratory, was compared with two other immunoassays, western blot (WB) and a sandwich immunochromatographic commercial kit (SI), and also with a direct polymerase chain reaction (PCR) method that detects proviral DNA. IFA showed to be a test with high sensitivity and specificity, and could be useful as a diagnostic tool in epidemiological studies. The presence of a low percentage of results with non-specific reactivity in the IFA could be resolved with further testing or use of an alternative method.

Molecular differentiation of Trichinella spiralis, T. pseudospiralis, T. papuae and T. zimbabwensis by pyrosequencing

2013 ◽  
Vol 89 (1) ◽  
pp. 118-123 ◽  
Author(s):  
L. Sadaow ◽  
C. Tantrawatpan ◽  
P.M. Intapan ◽  
V. Lulitanond ◽  
T. Boonmars ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Trichinella Spiralis ◽  
Large Subunit ◽  
Pcr Amplification ◽  
Epidemiological Studies ◽  
Chain Reaction ◽  
Target Region ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Zoonotic Parasites

AbstractNematodes of the genus Trichinella which infect wildlife and domestic animals show a cosmopolitan distribution. These zoonotic parasites are the aetiological agents of a severe human disease, trichinellosis. Twelve taxa are recognized in the Trichinella genus, but they cannot be identified by morphology since they are sibling species/genotypes. For epidemiological studies, it is extremely important to identify each taxon since they have different distribution areas and host ranges. In the present study, polymerase chain reaction (PCR) amplification of the mitochondrial large subunit ribosomal RNA (lsu-RNA) gene coupled with a pyrosequencing technique was developed to distinguish among four Trichinella species: Trichinella spiralis, T. pseudospiralis, T. papuae and T. zimbabwensis. A PCR method was used to amplify the lsu-RNA of Trichinella sp. larvae in mouse muscles and single larvae collected from infected muscles by digestion. The results show that the four Trichinella species can be distinguished by using 26 nucleotides in the target region and the method is sensitive enough to identify individual larvae. The pyrosequencing provides a simple, rapid and high-throughput tool for the differentiation of Trichinella species.

scholarly journals RECCURENT GIANT CONDYLOMATA ACUMINATA CAUSED BY HUMAN PAPILLOMA VIRUS IN HIV WITH HOMOSEXUAL MALE

2020 ◽  
Vol 8 (2) ◽  
pp. 131
Author(s):  
Emy Kusumaningsih ◽  
Lita Setyowatie
Keyword(s):  
Condylomata Acuminata ◽  
High Sensitivity ◽  
Surgical Excision ◽  
Papilloma Virus ◽  
Surgical Department ◽  
Lung Tuberculosis ◽  
Immunodeficiency Virus ◽  
Hpv Types ◽  
Polymerase Chain ◽  
Sex With Men

Perianal giant condylomata acuminate (GCA) is a rare clinical condition associated with low-risk Human papillomavirus (HPV) type 6 and 11 infections. Human Immunodeficiency Virus (HIV) infection is one of the risk factors for GCA, that can increase the condylomata acuminate incidence and spread caused by HPV. A 28-year-old man came with a cauliflower-like mass complaint in his perianal and anal since 2 months ago. The patient did not complain of pain or itching on the mass but often bled when defecating. The patient is a male who has sex with men (MSM) and often changes partners. He has been diagnosed with HIV since 11 months ago and regularly taking anti-retroviral drugs, Efavirenz 600 mg daily. He was also diagnosed having lung tuberculosis at the same time, got 6 months treatment and was declared cured. The venereological examination of the perianal and anal region revealed erythematous and grayish stem-shaped vegetation and papules, verrucous surface, multiple, well defined, with 3 x 1.5 x 2 cm in size. A positive act of white examination was obtained. Blood tests revealed CD+4 230 cells /μL. Polymerase chain reaction (PCR) examination for HPV obtained HPV types 6 and 11 infections. Histopathologic examination revealed acanthosis, papillomatosis, and hyperkeratotic epidermis and koilocytotic cells. The patient was treated with electrodesiccation three times but obtained mass in anal getting bigger with a size of 6 x 3 x 3 cm. Therefore, he agreed to be referred to the surgical department with an extensive surgical excision plan. Screening of GCA using PCR is not a routine examination but PCR has high sensitivity and specificity for determining the type of HPV, is useful for determining GCA prognosis and therapy, and is recommended for malignant and possible GCA recurrence detection

scholarly journals Anaplasma marginale infection in cattle from south-western Amazonia

2010 ◽  
Vol 30 (3) ◽  
pp. 249-254 ◽  
Author(s):  
Luciana Gatto Brito ◽  
Márcia Cristina de Sena Oliveira ◽  
Rodrigo Barros Rocha ◽  
Francelino Goulart da Silva Netto ◽  
Adriana Denise Marim ◽  
...  
Keyword(s):  
High Frequency ◽  
Simple Procedure ◽  
Anaplasma Marginale ◽  
Epidemiological Data ◽  
Epidemiological Studies ◽  
Chain Reaction ◽  
Blood Clots ◽  
Pcr Method ◽  
The Mean ◽  
Polymerase Chain

The present study provides the first epidemiological data regarding infection by Anaplasma marginale in cattle reared in south-western Brazilian Amazonia. One simple procedure was adapted for the extraction of DNA from blood clots collected in seven microregions of Rondônia State and two mesoregions of Acre State. PCR method was used to asses the frequency of A. marginale infections in 4 to12-month-old cattle. The cattle infection was investigated by polymerase chain reaction (PCR) using the specific primer "msp5" for A. marginale. The DNA amplifications revealed that the mean frequency of A. marginale infection was 98.6% (1,627/1,650) in samples from Rondonia, and 92.87% (208/225) in samples from Acre. The high frequency of A. marginale infections in 4 to 12-month-old cattle indicate a situation of enzootic stability in the studied areas and are comparable to those detected by immunodiagnosis in different endemic regions in Brazil. The DNA extraction of clotted blood method described here can be used for epidemiological studies on anaplasmosis and other bovine hemoparasites.

scholarly journals Use of Oropharyngeal Washes to Diagnose and Genotype Pneumocystis jirovecii

2015 ◽  
Vol 2 (3) ◽  
Author(s):  
Jonathan J. Juliano ◽  
Eric Barnett ◽  
Christian M. Parobek ◽  
Steve M. Taylor ◽  
Steven R. Meshnick ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
High Sensitivity ◽  
Diagnostic Techniques ◽  
Surface Glycoprotein ◽  
Pneumocystis Jirovecii ◽  
Major Surface Glycoprotein ◽  
Immunodeficiency Virus ◽  
Noninvasive Samples ◽  
Strain Type ◽  
Major Surface

Abstract Pneumocystis jirovecii is a symbiotic respiratory fungus that presents in 2 clinical forms: pneumonia in immunocompromised patients or colonization, defined by the presence of the organism without associated clinical symptoms. Currently, diagnosis requires invasive bronchoscopy, which may not be available in some settings and is inappropriate for detecting colonization in healthy individuals. Noninvasive diagnostic techniques and molecular strain typing tools that can be used on these samples are critical for conducting studies to better understand transmission. We evaluated 2 real-time polymerase chain reaction (PCR) assays targeting dihydropteroate synthase and the major surface glycoprotein for detection in 77 oropharyngeal washes (OPWs) from 43 symptomatic human immunodeficiency virus-infected patients who underwent bronchoscopy. We also evaluated the ability of a new microsatellite (MS) genotyping panel to strain type infections from these samples. Each PCR used individually provided a high sensitivity (>80%) for detection of pneumonia but a modest specificity (<70%). When used in combination, specificity was increased to 100% with a drop in sensitivity (74%). Concentration of organisms by PCR in the OPW tended to be lower in colonized individuals compared with those with pneumonia, but differences in concentration could not clearly define colonization in symptomatic individuals. Oropharyngeal wash samples were genotyped using 6 MSs with ≥4 alleles successfully genotyped in the majority of colonized patients and ≥5 alleles in patients with pneumonia. The MS profile was consistent over time within patients with serial OPWs analyzed. Microsatellite genotyping on noninvasive samples may aid in studying the molecular epidemiology of this pathogen without requiring invasive diagnostic techniques.

scholarly journals Detection and Monitoring of Greeneria uvicola and Colletotrichum acutatum Development on Grapevines by Real-Time PCR

Plant Disease ◽  
2011 ◽  
Vol 95 (3) ◽  
pp. 298-303 ◽  
Author(s):  
Suren K. Samuelian ◽  
Lindsay A. Greer ◽  
Sandra Savocchia ◽  
Christopher C. Steel
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Epidemiological Studies ◽  
Colletotrichum Acutatum ◽  
Internal Transcribed Spacer Region ◽  
Bitter Rot ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Species Specific ◽  
And Control

Bitter rot (Greeneria uvicola) and ripe rot (Colletotrichum acutatum, syn. C. simmondsii) occur frequently in subtropical grape-growing regions of Australia, where they cause yield loss and bitter taints in wine. To further advance the epidemiological studies of G. uvicola and C. acutatum and contribute toward their effective management and control, a rapid and reliable species-specific real-time polymerase chain reaction (PCR) method was developed based on the polymorphic portion of the internal transcribed spacer region of the two fungi. It was found that, within 6 to 8 h postinoculation, the assay could detect as little as 20 fg of genomic DNA and 10 conidia for both species. Artificially and naturally infected grape inflorescences and mature berries were analyzed by both conventional plating methods and real-time PCR. Fungal presence was demonstrated on all plant material but development was observed only on mature berries. The results demonstrate that the real-time PCR technique is a highly specific, rapid, and sensitive method that can be used to detect and study the dynamics of G. uvicola and C. acutatum during different stages of infection and on different grape tissues.

scholarly journals Natural Plasmodium infection in neotropical primates in the island of São Luís, state of Maranhão, Brazil

2015 ◽  
Vol 24 (2) ◽  
pp. 122-128 ◽  
Author(s):  
Mayra Araguaia Pereira Figueiredo ◽  
Silvia Maria Fátima Di Santi ◽  
Thaysa Araguaia Pereira Figueiredo ◽  
Rosangela Zacarias Machado
Keyword(s):  
Nonhuman Primates ◽  
Optical Microscope ◽  
Diagnostic Techniques ◽  
Immunofluorescence Assay ◽  
Wild Animal ◽  
Plasmodium Infection ◽  
Neotropical Primates ◽  
Chain Reaction ◽  
Free Living ◽  
Polymerase Chain

The states that make up the Legal Amazon Region, which include the state of Maranhão, account for 99% of registered cases of human malaria in Brazil. It is also believed that transmission of malaria from nonhuman primates (NHP) to humans occurs in this region, because of current reports of seroepidemiological results from samples from humans and NHP coexisting in the same areas. This study aimed to make morphological, serological and molecular diagnoses of Plasmodium spp. in neotropical primates on the island of São Luís, state of Maranhão, Brazil. The diagnostic techniques used were optical microscopy, the polymerase chain reaction (PCR) and the indirect immunofluorescence assay (IFA). From June 2009 to April 2010, 70 NHP were sampled: 50 at the Wild Animal Screening Center (CETAS), located in the municipality of São Luís and 20 free-living individuals that were caught in a private reserve located in the municipality of São Jose de Ribamar, state of Maranhão. Under an optical microscope, 140 slides (two from each animal) were evaluated and five animals (7.1%) were found to be positive. IFA did not detect anti-Plasmodium spp. From PCR on the 70 animals sampled, amplified Plasmodium spp. products were observed in 13 samples, of which eight (61.5%) were from free-living animals and five (38.5%) were from animals at CETAS.

scholarly journals Sensitivity and Specificity of a Monoclonal Antibody-Based Fluorescence Assay for Detecting Enterocytozoon bieneusi Spores in Feces of Simian Immunodeficiency Virus-Infected Macaques

2005 ◽  
Vol 12 (10) ◽  
pp. 1141-1144 ◽  
Author(s):  
Inderpal Singh ◽  
Abhineet S. Sheoran ◽  
Quanshun Zhang ◽  
Angela Carville ◽  
Saul Tzipori
Keyword(s):  
Monoclonal Antibody ◽  
Simian Immunodeficiency Virus ◽  
Chronic Diarrhea ◽  
Immunofluorescence Assay ◽  
Enterocytozoon Bieneusi ◽  
Diagnostic Assay ◽  
Specificity And Sensitivity ◽  
Immunodeficiency Virus ◽  
Pcr Method

ABSTRACT Enterocytozoon bieneusi is clinically the most significant among the microsporidia causing chronic diarrhea, wasting, and cholangitis in individuals with human immunodeficiency virus/AIDS. Microscopy with either calcofluor or modified trichrome stains is the standard diagnostic test for microsporidiosis and does not allow species identification. Detection of E. bieneusi infection based on PCR is limited to a few reference laboratories, and thus it is not the standard diagnostic assay. We have recently reported the development and characterization of a panel of monoclonal antibodies against E. bieneusi, and in this publication we evaluated the specificity and sensitivity of an immunofluorescence assay (IFA), compared with PCR, in simian immunodeficiency virus-infected macaques. The IFA, which correlated with the primary PCR method, with a detection limit of 1.5 ×105 spores per gram of feces, will simplify considerably the detection of E. bieneusi spores in clinical and environmental specimens and in laboratory and epidemiological investigations.

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