Improvement of a PCR test to diagnose infection by Mansonella ozzardi (original) (raw)
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Parasites & Vectors, 2015
Background: Mansonella ozzardi is a poorly understood human filarial parasite with a broad distribution throughout Latin America. Most of what is known about its parasitism has come from epidemiological studies that have estimated parasite incidence using light microscopy. Light microscopy can, however, miss lighter, submicroscopic, infections. In this study we have compared M. ozzardi incidence estimates made using light microscopy, with estimates made using PCR. Methods: 214 DNA extracts made from Large Volume Venous Blood Samples (LVVBS) were taken from volunteers from two study sites in the Rio Solimões region: Codajás [n = 109] and Tefé [n = 105] and were subsequently assayed for M. ozzardi parasitism using a diagnostic PCR (Mo-dPCR). Peripheral finger-prick blood samples were taken from the same individuals and used for microscopic examination. Finger-prick blood, taken from individuals from Tefé, was also used for the creation of FTA®card dried blood spots (DBS) that were subsequently subjected to Mo-dPCR. Results: Overall M. ozzardi incidence estimates made with LVVBS PCRs were 1.8 times higher than those made using microscopy (44.9 % [96/214] compared with 24.3 % [52/214]) and 1.5 times higher than the PCR estimates made from FTA®card DBS (48/105 versus 31/105). PCR-based detection of FTA®card DBS proved 1.3 times more sensitive at diagnosing infections from peripheral blood samples than light microscopy did: detecting 24/105 compared with 31/105. PCR of LVVBS reported the fewest number of false negatives, detecting: 44 of 52 (84.6 %) individuals diagnosed by microscopy; 27 of 31 (87.1 %) of those diagnosed positive from DBSs and 17 out of 18 (94.4 %) of those diagnosed as positive by both alternative methodologies. Conclusions: In this study, Mo-dPCR of LVVBS was by far the most sensitive method of detecting M. ozzardi infections and detected submicroscopic infections. Mo-dPCR FTA®card DBS also provided a more sensitive test for M. ozzardi diagnosis than light microscopy based diagnosis did and thus in settings where only finger-prick assays can be carried-out, it may be a more reliable method of detection. Most existing M. ozzardi incidence estimates, which are often based on light microscope diagnosis, are likely to dramatically underestimate true M. ozzardi parasitism incidence levels.
Memorias do Instituto Oswaldo Cruz, 2018
The human filarial worm Mansonella ozzardi is highly endemic in the large tributaries of the Amazon River. This infection is still highly neglected and can be falsely negative when microfilariae levels are low. This study investigated the frequency of individuals with M. ozzardi in riverine communities in Coari municipality, Brazilian Amazon. Different diagnostic methods including polymerase chain reaction (PCR), blood polycarbonate membrane filtration (PCMF), Knott's method (Knott), digital thick blood smears (DTBS) and venous thick blood smears (VTBS) were used to compare sensitivity and specificity among the methods. Data were analysed using PCMF and Bayesian latent class models (BLCM) as the gold standard. We used BLCM to calculate the prevalence of mansonelliasis based on the results of five diagnostic methods. The prevalence of mansonelliasis was 35.4% by PCMF and 30.1% by BLCM. PCR and Knott methods both possessed high sensitivity. Sensitivity relative to PCMF was 98.5% [...
Memórias do Instituto Oswaldo Cruz, 2014
The prevalence of mansonelliasis was studied in the municipality of Tefé, state of Amazonas, Brazil. The prevalence (thick blood smear method) was 13.6% (147/1,078), higher in the Solimões River region (16.3%) than in the Tefé River region (6.3%). In the sampled communities in the Solimões River region, a higher density of cases was observed, as indicated by a kernel analysis (odds ratio 0.34; 95% confidence interval: 0.20-0.57). Males had a higher prevalence (χ2 = 31.292, p < 0.001) than women. Mansonella ozzardi prevalence was higher in retirees and farmers (28.9% and 27%, respectively). Prevalence also significantly increased with age (χ2 = -128.17, p < 0.001), with the highest numbers occurring in persons older than 67 years.
Memórias do Instituto Oswaldo Cruz, 2014
Rivers in the state of Amazonas (AM). However, little is known about the presence of this parasite in communities located in regions bordering AM and the state of Acre. The prevalence rate of M. ozzardi infections was determined in blood samples from volunteers according to the Knott method. A total of 355 volunteers from six riverine communities were enrolled in the study and 65 (18.3%) were found to be infected with M. ozzardi. As expected, most of the infections (25%) occurred in individuals involved in agriculture, cattle rearing and fishing and an age/sex group analysis revealed that the prevalence increased beginning in the 40-50-years-of-age group and reached 33% in both sexes in individuals over 50 years of age. Based on the described symptomatology, articular pain and headache were found to be significantly higher among infected individuals (56 and 65% prevalence, respectively, p < 0.05). Sera from volunteers were subjected to ELISA using a cocktail of recombinant proteins from Onchocerca volvulus to evaluate the specificity of the test in an endemic M. ozzardi region. No cross-reactions between M. ozzardi-infected individuals and recombinant O. volvulus proteins were detected, thus providing information on the secure use of this particular cocktail in areas where these parasites are sympatric.
Atypical Mansonella ozzardi Microfilariae from an Endemic Area of Brazilian Amazonia
The American journal of tropical medicine and hygiene, 2016
Mansonellosis is endemic in several regions of Africa, the Caribbean, and Latin America. Mansonella ozzardi and Mansonella perstans have been reported in Latin America, including the Amazon region. A morphological and molecular microfilariae study was performed in Pauini (Brazil). Blood samples were collected from 40 individuals, and were analyzed by Giemsa-stained blood film and by two different nested polymerase chain reactions which detect internal transcribed spacer-1 and the major sperm protein gene. By microscopy, 14 of 40 were positive: 11 as M. ozzardi and three as M. perstans-like infections. Both molecular methods detected 19 positive cases as M. ozzardi, including those 14 individuals detected by microscopy, without detectable genetic differences among any of the 19 positive samples. Molecular techniques showed an improvement of mansonellosis diagnosis and may become an effective tool to evaluate the present status of M. ozzardi and M. perstans in Latin America.
Transfusion, 2018
BACKGROUNDIn the Brazilian Amazon, the filarial nematode Mansonella ozzardi co‐exists with malaria parasites and thick blood smear microscopy is considered the diagnostic gold standard. Transfusion of M. ozzardi microfilariae does not establish new infections, however microfilariae can survive approximately 2 years in blood‐recipients with unknown risk of pathology. Data on transfusion‐transmitted filariasis are lacking. This study investigated M. ozzardi parasitemias in blood donors from decentralized centers of “Fundação Hematologia e Hemoterapia do Estado do Amazonas/HEMOAM,” Northern Brazil.STUDY DESIGN AND METHODSCross‐sectional investigation employing blood smear microscopy (n = 356) and qualitative nested‐M. ozzardi‐PCR (227 out of 356) in donor candidates from 19 hemocenters in interior/rural municipalities of Amazonas state. FINDINGS: Participants were mostly young males. Positivity by microscopy was 7.9% (28 out of 356) and 23.8% by M. ozzardi‐PCR (54 out of 227). Parasita...
Revista da Sociedade Brasileira de Medicina Tropical, 2016
Introduction: Mansonella ozzardi is a widely distributed fi laria worm in the Amazon region. This study aimed to determine the prevalence of M. ozzardi infection in riverine communities of Lábrea municipality, Amazonas State, Brazil. Methods: A diagnostic blood fi ltration method in a polycarbonate membrane was used. Results: M. ozzardi was found in 50.3% of the sample, with the highest prevalence in farmers/fi shermen (69.4%; χ 2 = -19.14, p<0.001). The prevalence was higher in longerterm residents (≥11 years; 60.2%). Conclusions: M. ozzardi infection rates are high near the Purus River, much greater than those previously reported based on diagnosis using thick blood smears.
Memórias do Instituto Oswaldo Cruz, 2010
We present filaria-nested polymerase chain reaction (PCR), which is based on amplification of first internal transcribed spacer rDNA to distinguish three parasitic filarial species (Onchocerca volvulus, Mansonella ozzardi and Mansonella perstans) that can be found in the Amazon Region. Nested PCR-based identifications yielded the same results as those utilizing morphological characters. Nested PCR is highly sensitive and specific and it detects low-level infections in both humans and vectors. No cross-amplifications were observed with various other blood parasites and no false-positive results were obtained with the nested PCR. The method works efficiently with wholeblood, blood-spot and skin biopsy samples. Our method may thus be suitable for assessing the efficacy of filaria control programmes in Amazonia by recording parasite infections in both the human host and the vector. By specifically differentiating the major sympatric species of filaria, this technique could also enhance epidemiological research in the region.
High Prevalence of Mansonella ozzardi Infection in the Amazon Region, Ecuador
Emerging Infectious Diseases, 2019
We reviewed Giemsa-stained thick blood smears, obtained through the national malaria surveillance program in the Amazon region of Ecuador, by light microscopy for Mansonella spp. microfilariae. Of 2,756 slides examined, 566 (20.5%) were positive. Nested PCR confirmed that the microfilariae were those of M. ozzardi nematodes, indicating that this parasite is endemic to this region. A lthough mansonelliases is probably the most prevalent filarial infection worldwide, it is the least studied and is considered a neglected parasite infection (1,2). Human infection with members of the filarial nematode genus Mansonella, including M. ozzardi, M. perstans, and M. streptocerca nematodes, is common and widespread in the Western Hemisphere and Africa. M. ozzardi nematodes are found exclusively in the Western Hemisphere from southern Mexico to northwestern Argentina but have not been reported in Ecuador, Chile, Uruguay, and Paraguay (1,2). M. perstans infections are found mainly in sub-Saharan Africa, with sporadic cases in a few countries in South America, whereas M. streptocerca infections are found only in Africa (1-4). Epidemiologic studies have reported that M. ozzardi nematodes are highly prevalent in the Amazon Basin (Brazil, Colombia, Peru, Bolivia, Venezuela, and Argentina) and on some Caribbean islands. In the general population, the prevalence rate of infection ranges from 0% to 46%; however, in some areas it is 92.3% (2). Community health workers from the malaria control program in the Amazon region of Ecuador have reported that filaria-like nematodes are seen in thick blood smears, suggesting the presence of a Mansonella sp. in this area. Mansonella spp. nematodes are transmitted by dipteran flies. In particular, M. ozzardi nematodes are transmitted by biting midges of the genera Culicoides and by black flies from the genus Simulium (1). Both of these vectors are present in the Amazon region of Ecuador (Renato Leon, Universidad San Francisco de Quito, pers. comm., 2018 Dec 5).
Revista da Sociedade Brasileira de Medicina Tropical, 2011
Introduction: Mansonella ozzardi, a human filarial nematode transmitted by Diptera Simuliidae flies (black flies), is widely spread in the Amazon, while the occurrence of mansonelliasis in the State of Rondônia (Western Amazonia) has not been reported thus far. The existence of this parasitic disease in this state is investigated in this study. Methods: A total of 4,452 people who lived alongside the Madeira, Mamore, Guapore, Machado and Preto Rivers, in the State of Rondônia, were examined using the thick blood smear method. Alongside the Machado and Guapore rivers, 2,112 adult simuliid specimens were captured. Results: Infected people were not reported, although simuliids with vector potential such as Cerqueirellum pydanielli, C. argentiscutum and C. amazonicum were found. Conclusions: This study registered no infected people. This finding may be explained by the individuals' low level of microfilaremia that is below the limit of detection of the diagnostic method used in the study or insufficient migration of people infected with M. ozzardi from endemic areas, making it impossible to introduce the disease into Rondônia. Further studies using more sensitive diagnostic techniques could shed light on this question.