scholarly journals Leishmania infantum infection in dogs from maroon communities in the Eastern Amazon

2017 ◽  
Vol 47 (1) ◽  
Author(s):  
Andréia Ferreira da Silva ◽  
Ágatha Rossanni Alves Damasceno ◽  
Welton Seabra Prado ◽  
Rossela Damasceno Caldeira ◽  
Francisco Dantas Sampaio-Junior ◽  
...  
Keyword(s):  
Canine Visceral Leishmaniasis ◽  
Risk Map ◽  
Intact Skin ◽  
Chain Reaction ◽  
Blood Samples ◽  
Factors Associated ◽  
Forested Areas ◽  
Polymerase Chain ◽  
Collection Sites ◽  
Skin Samples

ABSTRACT: This study was designed to detect L. infantum infection in dogs and to evaluate the factors associated with canine visceral leishmaniasis in the maroon communities of Menino Jesus de Petimandeua and Itaboca in the municipality of Inhangapi, Pará, Brazil. Whole blood and intact skin samples were collected from 143 dogs, and a questionnaire was applied. L. infantum DNA was detected by polymerase chain reaction (PCR) using primers RV1 and RV2. Collection sites were georeferenced to obtain a spatial distribution of the residences visited and infected dogs. L. infantum DNA was detected in 8.4% (12/143) of the skin samples and in 1.4% (2/143) of the blood samples. On the risk map, three clusters were observed in Itaboca and one was observed in Menino Jesus de Petimandeua. We observed that most of the inhabitants in these maroon communities live close to forested areas and do not use protection against insect vectors. The presence of canine reservoirs of L. infantum associated to environment characteristics (preserved forests and deforested areas) and habits of dog owners (living near forested areas and not using any protection against insects) may favor the transmission of L. infantum in the studied areas.

Comparison of Droplet Digital Polymerase Chain Reaction (ddPCR) and Real-Time Quantitative Polymerase Chain Reaction (qPCR) in Detecting Neonatal Invasive Fungal Infections

2021 ◽  
Vol 11 (3) ◽  
pp. 373-379
Author(s):  
Huitao Li ◽  
Xueyu Chen ◽  
Xiaomei Qiu ◽  
Weimin Huang ◽  
Chuanzhong Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Fungal Isolation ◽  
Blood Samples ◽  
Fungal Strains ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Invasive fungal infection (IFI) is the leading cause of death in neonatal patients, yet the diagnosis of IFI remains a major challenge. At present, most IFI laboratory diagnostic methods are based on classical, but limited, methods such as fungal isolation and culture and histopathological examination. Recently, quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) technology have been adopted to quantify nucleic-acid identification. In this study, we established qPCR and ddPCR assays for IFI diagnosis and quantification. qPCR and ddPCR were carried out using identical primers and probe for the amplification of 18S rRNA. Assay results for three fungal strains were positive, whereas ten non-fungal strains had negative results, indicating 100% specificity for both ddPCR and qPCR methods. Genomic DNA of Candida albicans was tested after a serial dilution to compare the sensitivity of the two PCR methods. The limit of detection of ddPCR was 3.2 copies/L, which was a ten-fold increase compared with that of the qPCR method (32 copies/L). Blood samples from 127 patients with high-risk factors and clinical symptoms for IFI were collected from a NICU in Shenzhen, China, and analyzed using qPCR and ddPCR. Thirty-four blood samples from neonates had a proven or probable diagnosis of IFI, and 25 of these were positive by qPCR, whereas 30 were positive by ddPCR. Among the 93 blood samples from neonates who had a possible IFI or no IFI, 24 were positive using qPCR, and 7 were positive using ddPCR. In conclusion, ddPCR is a rapid and accurate pan-fungal detection method and provides a promising prospect for IFI clinical screening.

scholarly journals Bartonella spp. in human and animal populations in Gauteng, South Africa, from 2007 to 2009

2012 ◽  
Vol 79 (2) ◽  
Author(s):  
Anastasia N. Trataris ◽  
Jennifer Rossouw ◽  
Lorraine Arntzen ◽  
Allan Karstaedt ◽  
John Frean
Keyword(s):  
Health Care Professionals ◽  
Deoxyribonucleic Acid ◽  
Bartonella Henselae ◽  
Hiv Positive ◽  
Host Immune System ◽  
Wild Rodent ◽  
Chain Reaction ◽  
Blood Samples ◽  
Animal Populations ◽  
Polymerase Chain

Bartonellae are highly adaptive organisms that have the ability to evade the host immune system and cause persistent bacteraemia by occupying the host’s erythrocytes. Bartonella spp. is under-studied and health care professionals often misdiagnose Bartonella-related infections. The aim of this study was to investigate the carriage of Bartonella spp. circulating in human and animal populations in Gauteng using culturing and polymerase chain reaction (PCR) detection. A total of 424 human, 98 cat, 179 dog, and 124 wild rodent blood samples were plated onto specialised media and incubated for 7–21 days at 37 ºC in CO2. Culture isolates morphologically similar to Bartonella control strains were confirmed by PCR and sequenced to determine species. Deoxyribonucleic acid (DNA) was extracted from all blood samples and tested by nested PCR. Bartonella could only be cultured from the cat and rodent specimens. Cat isolates were > 99% similar to Bartonella henselae URBHLIE 9, previously isolated from an endocarditis patient, and rat isolates were > 98% similar to either RN24BJ (candidus ‘Bartonella thailandensis’) or RN28BJ, previously isolated from rodents in China. The PCR prevalences were 22.5% in HIV-positive patients, 9.5% in clinically healthy volunteers, 23.5% in cats, 9% in dogs and 25% in rodents. Findings of this study have important implications for HIV-positive patients.

scholarly journals Evaluation on the presence of Anaplasma, Ehrlichia, and Babesia spp. in goats (Capra hircus) in Cebu, the Philippines

2019 ◽  
Vol 12 (6) ◽  
pp. 774-777 ◽  
Author(s):  
Adrian P. Ybañez ◽  
Orgil V. Arrabis ◽  
Dennis Justin M. Alvarez ◽  
Eloiza May S. Galon ◽  
Rhea Mae P. Jayag ◽  
...  
Keyword(s):  
Blood Smear ◽  
Peripheral Blood Smear ◽  
The Philippines ◽  
Capra Hircus ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Blood Samples ◽  
Blood Smear Examination ◽  
Polymerase Chain ◽  
Babesia Spp

Background: Tick-borne diseases are caused by a wide variety of viruses, pathogens, and diseases. Anaplasma, Ehrlichia, and Babesia spp. are among the most known tick-borne pathogens in Asia. In the Philippines, these pathogens were already reportedly present in dogs and large ruminants, but no study has been reported yet evaluating their presence in goats. Aim: The present study aimed to evaluate the presence of Anaplasma, Ehrlichia, and Babesia spp. in goats in Cebu, the Philippines. Materials and Methods: A total of 100 blood samples from goats were collected in Cebu, the Philippines. Profile of sampled goats including age, body score, and sex was obtained. Peripheral blood smear examination and DNA extraction were performed. Nested polymerase chain reaction assay was used to evaluate the presence of Anaplasma, Ehrlichia, and Babesia spp. Results: None of the samples were found positive with Anaplasma, Ehrlichia, and Babesia spp. infection. Conclusion: Tested goats were negative with Anaplasma, Ehrlichia, and Babesia spp. and calls for continuous surveillance of these pathogens due to the reported detection of these pathogens in other livestock animals in the area.

scholarly journals AMPLIFIKASI GEN COI DARI SAMPEL DARAH ULAR DENGAN MENGGUNAKAN BEBERAPA PASANGAN PRIMER UNIVERSAL

ZOOTEC ◽  
2019 ◽  
Vol 39 (2) ◽  
pp. 314
Author(s):  
Dylan R. Pahlevi ◽  
Edwin De Queljoe ◽  
Beivy J. Kolondam
Keyword(s):  
Coi Gene ◽  
Polymerase Chain Reaction Reaction ◽  
Cytochrome Oxidase Subunit ◽  
Chain Reaction ◽  
Dna Ladder ◽  
Blood Samples ◽  
Total Dna ◽  
Polymerase Chain ◽  
Snake Blood ◽  
I Gene

AMPLIFICATION OF COI GENE FROM SNAKE BLOOD SAMPLES USING TWO UNIVERSALPRIMER PAIRS. This study aims to amplify COI (Cytochrome Oxidase Subunit I) gene fragments from snake blood. Four samples were obtained from four different snake individuals that were captured in Tapahan Telu Waterfall, Kali Village, Minahasa Regency. Total DNA from the sample was isolated and then the COI gene was amplified through the PCR (Polymerase Chain Reaction) reaction using two primer pairs, LCO1490-HCO2198 and FF2d-FR1d. These four samples were successfully amplified using different primers, i.e. DRP1 and DRP3 by FF2d-FR1d and DRP2 and DRP4 by LCO1490-HCO2198 primers. The success of amplification marked by the presence of 710 bp (LCO1490-HCO2198) and 707 bp (FF2d-FR1d) bands, which were indicated by those bands were located close to the standard 750 bp DNA ladder. Key words: Snake, Amplification, COI gene, Primers, PCR

scholarly journals Factors Associated with HIV prevalence among HIV-exposed Infants with HIV DNA Polymerase Chain Reaction Tests in Malawi: 2013-2020

2021 ◽  
Author(s):  
Wingston Ng'ambi ◽  
Janne Estill ◽  
Fatma Aziza Merzouki ◽  
Erol Orel ◽  
Tiwonge Chimpandule ◽  
...  
Keyword(s):  
Hiv Infection ◽  
Temporal Trends ◽  
Chain Reaction ◽  
Factors Associated ◽  
Hiv Dna ◽  
Polymerase Chain ◽  
Hiv Exposed ◽  
Spatial And Temporal Trends ◽  
Dna Pcr ◽  
Test Result

Background: Despite the high availability of individual-level data of infants accessing HIV DNA polymerase chain reaction (DNA-PCR) testing service, there has been little in-depth analysis of such data. Therefore, we describe spatial and temporal trends in risk of HIV infection among Malawi HIV-exposed infants (HEI) with DNA-PCR HIV test result from 2013 to 2020. Methods: This is an implementation study using routinely collected patient-level HIV DNA-PCR test result data extracted from the national Laboratory Management Information System database managed by the Department of HIV/AIDS between 1 January 2013 and 30 June 2020. We calculated frequencies, proportions and odds ratios (OR) with their associated 95% confidence intervals (95%CI). We performed a random-effects logistic regression to determine the risk factors associated with HIV infection in infants, controlling for the spatial autocorrelation between districts and adjusting for other variables. Results: We evaluated 255,229 HEI across 750 facilities in 28 districts. The overall risk of HIV infection among all tested HEI between 2013 and 2020 was 7.2% (95%CI: 7.1-7.3). We observed a decreasing trend in the proportion of HEI that tested HIV positive from 7.0% (95%CI: 6.6-7.4) in 2013 to 5.7% (95%CI: 5.4-5.9) in 2015 followed by an increase to 9.9% (95%CI: 9.6-10.2) in 2017 and then a decreasing trend to 4.2% (95%CI: 3.7-4.6) in 2020. The risk of HIV infection increased by age of the HEI. There was spatial heterogeneity of HIV prevalence between districts of Malawi. Conclusion: We summarised spatial and temporal trends of risk of HIV infection amongst HEI in Malawi between 2013 and 2020. There is need for further strengthening of EID program to ensure that all the HEI are enrolled in care by eight weeks of age in order to further reduce mother-to-child transmission of HIV.

scholarly journals Molecular detection and occurrence of equine theileriosis in Arabian horses in Al-Najaf province/Iraq

2020 ◽  
Vol 57 (3) ◽  
pp. e166996
Author(s):  
Hayder Mohammad Al-Rammahi ◽  
Abdulameer Abed Hatem ◽  
Asaad Chasib Al-Atabi
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Molecular Detection ◽  
Molecular Technique ◽  
Chain Reaction ◽  
Blood Samples ◽  
Equine Piroplasmosis ◽  
Polymerase Chain

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the variations in the occurrences of equine piroplasmosis according to gender, age, and geographical areas.

scholarly journals Molecular identification by multiplex polymerase chain reaction of Pasteurella multocida in cattle and buffaloes in Baghdad

2014 ◽  
Vol 38 (1) ◽  
pp. 99-106
Author(s):  
Ihab G. M. AL-Shemmari
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Multiplex Polymerase Chain Reaction ◽  
Pasteurella Multocida ◽  
Type B ◽  
Chain Reaction ◽  
Blood Samples ◽  
Nasal Swabs ◽  
Polymerase Chain ◽  
B 31

The aim of this study was to identify pasteurella multocida and their types by PCR in cattle’s and buffaloesi bagdad from March to August 2012 on 204 animals , including 102 cattle and 102 buffaloes at slaughter houses from Baghdad .Blood samples and nasal swaps were collected , before slaughtering and lung tissues of slaughtered animal , and from 54 clinically suspected cases of pasteurellosis , including 27 bovines ,and 27 buffaloes the samples taken included blood and nasal swabs . Pasteurellamultocida were isolated from 94 animals include 49 cattle 45 buffaloes. The typing of the isolates by multiplex PCR for genotyping Pasteuerllamultocida revealed 93 isolates of type B , 31 from cattle and 62 from buffaloes ,and 81 isolates of type A , 55 from cattle and 26 from buffaloes .

Prognostic Value of Circulating Melanoma Cells Detected by Reverse Transcriptase–Polymerase Chain Reaction

2003 ◽  
Vol 21 (5) ◽  
pp. 767-773 ◽  
Author(s):  
Giuseppe Palmieri ◽  
Paolo A. Ascierto ◽  
Francesco Perrone ◽  
Sabrina M.R. Satriano ◽  
Alessandro Ottaiano ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Subgroup Analysis ◽  
Peripheral Blood ◽  
Predictive Value ◽  
Prognostic Value ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Blood Samples ◽  
Stage Of Disease ◽  
Polymerase Chain

Purpose: Factors that are predictive of prognosis in patients who are diagnosed with malignant melanoma (MM) are widely awaited. Detection of circulating melanoma cells (CMCs) by reverse transcriptase-polymerase chain reaction (RT-PCR) has recently been postulated as a possible negative prognostic factor. Two main questions were addressed: first, whether the presence of CMCs, defined as the patient being positive for any of the three markers, had a prognostic role; and second, what the predictive value of each individual marker was. Patients and Methods: A consecutive series of 200 melanoma patients observed between January 1997 and December 1997, with stage of disease ranging from I to IV, was analyzed by semiquantitative RT-PCR. Tyrosinase, p97, and MelanA/MART1 were used as markers to CMCs on baseline peripheral blood samples. Progression-free survival (PFS) was used as a unique end point and was described by the product limit method. Multivariable analysis was applied to verify whether the auspicated prognostic value of these markers was independent of the stage of disease, and a subgroup analysis was performed that excluded patients with stage IV disease. Results: Overall, 32% (64 of 200) of patients progressed, and a median PFS of 52 months in the whole series was observed. The presence of CMCs and the markers individually or combined was predictive of prognosis in the univariate analysis but did not provide additional prognostic information to the stage of disease in multivariable models. In the subgroup analysis of stage (ie, I–III subgroup), similar results were observed. Conclusion: Detection of CMCs in peripheral blood samples at the time of MM diagnosis by semiquantitative RT-PCR does not add any significant predictive value to the stage of disease. Thus, this approach should not be used in clinical practice, and further studies are required to determine its usefulness.

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