Comparison of serology, antigenemia assay and the polymerase chain reaction for monitoring active cytomegalovirus infections in hematopoietic stem cell transplantation patients

Forty-six allogeneic hematopoietic stem cell transplantation (HSCT) patients were monitored for the presence of CMV antibodies, CMV-DNA and CMV antigens after transplantation. Immunoenzymatic serological tests were used to detect IgM and the increase in CMV IgG antibodies (increase IgG), a nested polymerase chain reaction (N-PCR) was used to detect CMV-DNA, and an antigenemia assay (AGM) was used to detect CMV antigens. The presence of CMV-IgM and/or CMV-increase IgG antibodies was detected in 12/46 (26.1%) patients, with a median time between HSCT and the detection of positive serology of 81.5 days. A positive AGM was detected in 24/46 (52.2%) patients, with a median time between HSCT and antigen detection of 62 days. Two or more consecutive positive N-PCR results were detected in 32/46 (69.5%) patients, with a median time between HSCT and the first positive PCR of 50.5 days. These results confirmed that AGM and mainly PCR are superior to serology for the early diagnosis of CMV infection. Six patients had CMV-IgM and/or CMV-increase IgG with a negative AGM (five cases) or N-PCR assay (one case). In five of these cases the serological markers were detected during the first 100 days after HSCT, the period of highest risk. These findings support the idea that serology may be useful for monitoring CMV infections in HSCT patients, especially when PCR is unavailable.

Survival of Trypanosoma cruzi in sugar cane used to prepare juice

Chagas disease can be transmitted to man by many different means, including contact with infected triatomine feces, blood transfusion, laboratory accidents, organ transplants, and congenital or oral routes. The latter mode has received considerable attention recently. In this assay, we evaluate the survival of Trypanosoma cruzi contaminating sugar cane used to prepare juice, as well as the viability and capacity for infection by the parasite after recovery. Thirty triatomines were contaminated with T. cruzi Y strain and 45 days later pieces of sugar cane were contaminated with the intestinal contents of the insects. The pieces were ground at different intervals after contamination (time = 0, 1, 4, 6, 12 and 24 hours) and the juice extracted and analyzed. Different methods were used to show T. cruzi in the juice: direct analysis, hematocrit tube centrifugation and QBC, and experimental inoculation in 47 female BALB/c mice (five control mice and seven mice for each interval examined (five inoculated orally and two intraperitoneally). Positive results were found using the direct analysis and QBC methods for juice prepared up to 12 hours after initial contamination. However, by the centrifugation technique, positivity was found only up to four hours after contamination of the sugar cane. Inoculated animals showed parasitemia during a 14 day observation period, demonstrating the high survival rate of T. cruzi in sugar cane.

Serodiagnosis of neurocysticercosis in patients with epileptic seizure using ELISA and immunoblot assay

Sera from 88 patients from Santa Catarina and São Paulo states of Brazil, with epileptic seizures who underwent cerebral computed tomography (CT) were analyzed for the detection of antibodies to T. solium cysticercus by ELISA and Immunoblot (IB) with the following antigens: Taenia solium cysticercus total saline (Tso), Taenia crassiceps cysticercus vesicular fluid (Tcra-vf) and T. crassiceps cysticercus glycoproteins (Tcra-gp). ELISA carried out with Tso, Tcra-vf and Tcra-gp antigens showed 95%, 90% and 80% sensitivities, respectively, and 68%, 85% and 93% specificities, respectively. In the epileptic patients group, ELISA positivity was 30%, 51% and 35% with Tso, Tcra-vf and Tcra-gp antigens respectively. Considering the IB as the confirmatory test, the positivity was 16% (14/88) in the epileptic patients total group and 22% (12/54) in the epileptic patients with positive CT and signals of cysticercosis. We found a significant statistical correlation among ELISA or IB results and the phase of the disease when any antigens were used (p < 0.05). We emphasize the need to introduce in the laboratory routine the search for neurocysticercosis (NC) in patients presenting with epileptic seizures because of the high risk of acquiring NC in our region and its potential cause of epilepsy.

Giardia lamblia and other intestinal parasitic infections and their relationships with nutritional status in children in Brazilian Amazon

The objective of this survey was to assess the relationships between intestinal parasitism, nutritional status and hemoglobin level in children with Indian ascendancy living in an urban area in Brazilian Amazon. We carried out a cross-sectional survey obtaining anthropometric, parasitological and socioeconomic data, and hemoglobin measurements of children aged six to 84 months. Anthropometric data were expressed as z-scores for weight for age (WAZ), height for age (HAZ), weight for height (WHZ) and mid upper circumference for age (MUACZ) parameters. Parasitological examinations were performed through Ritchie (n = 307), Kato-Katz (n = 278), Baermann-Moraes (n = 238) and Safranin-methylene blue methods (n = 307). Hemoglobin measurements were obtained with a Hemocue® photometer (n = 282). Socioeconomic data were used in order to classify children in three family income strata (n = 242). Multiple linear regression analysis showed independent interactions between Giardia lamblia and WAZ (beta = -0.195, SE = 0.138, p = 0.003), WHZ (beta = -0.161, SE = 0.133, p = 0.018) and MUACZ (beta = -0.197, SE = 0.143, p = 0.011), controlling for age, sex, family income, Ascaris lumbricoides, and hookworm infection. Also, the multivariate model showed that the only variable associated with hemoglobin levels was age. Intestinal parasitism control should increase children's possibilities of full development in the studied area.

Intestinal parasitoses and environmental factors in a rural population of Argentina, 2002-2003

We evaluate the prevalence of intestinal parasites in 504 people and the degree of association between environmental variables and parasites found in population, soil and water in a rural area of Argentina during 2002-2003. A structured survey was used to evaluate the environmental variables and fecal-human, soil and water samples were analyzed. The prevalence of parasites was 45.4%. Most prevalent protozoa were Blastocystis hominis (27.2%) and Giardia lamblia (6.9%), while the most prevalent helminth was Ascaris lumbricoides (3.8%). The analyzed environmental variables showing association (p < 0.05) with presence of parasites in population were: cardboard-tin or wooden house, dirt floor, home or communal water pump, faucet outside the house or public faucet and cesspool or latrine. Parasite forms were found in 82.3% of the soil samples and in 84.2% of the water samples. In both samples we found parasites that were also found in people. In this study we have found deficient sanitary conditions associated with presence of parasites in population and we have evidenced that contaminated soil and water were the source of these parasites.

Prevalence of intestinal microsporidiosis in Human Immunodeficiency Virus-infected patients with diarrhea in major United States cities

To determine the prevalence of intestinal microsporidiosis in HIV-infected patients, we performed a prospective study of HIV-infected patients with diarrheal illnesses in three US hospitals and examined an observational database of HIV-infected patients in 10 US cities. Among 737 specimens from the three hospitals, results were positive for 11 (prevalence 1.5%); seven (64%) acquired HIV through male-to-male sexual contact, two (18%) through male-to-male sexual contact and injection drug use, and one (9%) through heterosexual contact; one (9%) had an undetermined mode of transmission. Median CD4 count within six months of diagnosis of microsporidiosis was 33 cells/µL (range 3 to 319 cells/µL). For the national observational database (n = 24,098), the overall prevalence of microsporidiosis was 0.16%. Prevalence of microsporidiosis among HIV-infected patients with diarrheal disease is low, and microsporidiosis is most often diagnosed in patients with very low CD4+ cell counts. Testing for microsporidia appears to be indicated, especially for patients with very low CD4+ cell counts.

Prevalence of intestinal parasites in children from public daycare centers in the city of Belo Horizonte, Minas Gerais, Brazil

The objective of this study was to verify the occurrence of intestinal parasites in 3 to 6-year-old children from daycare centers maintained by the municipal government of Belo Horizonte, Minas Gerais, Brazil. Coproparasitological tests performed in 472 children have shown that 24.6% of them had some type of parasites, 6.6% of the children having more than one type. Among protozoa, Entamoeba coli (14.0%) and G. duodenalis (9.5%) were the most prevalent, whereas Ascaris lumbricoides (3.0%) and Trichuris trichiura (1.1%) were the most frequent among the helminths. Thus, we can observe that intestinal parasites still represent a serious public health problem in Belo Horizonte, especially among children and in areas where the socioeconomic conditions are less favorable.

Molecular characterization of two rocio flavivirus strains isolated during the encephalitis epidemic in são paulo state, brazil and the development of a one-step rt-pcr assay for diagnosis

Rocio virus (ROCV) was responsible for an explosive encephalitis epidemic in the 1970s affecting about 1,000 residents of 20 coastland counties in São Paulo State, Brazil. ROCV was first isolated in 1975 from the cerebellum of a fatal human case of encephalitis. Clinical manifestations of the illness are similar to those described for St. Louis encephalitis. ROCV shows intense antigenic cross-reactivity with Japanese encephalitis complex (JEC) viruses, particularly with Ilheus (ILHV), St. Louis encephalitis, Murray Valley and West Nile viruses. In this study, we report a specific RT-PCR assay for ROCV diagnosis and the molecular characterization of the SPAn37630 and SPH37623 strains. Partial nucleotide sequences of NS5 and E genes determined from both strains were used in phylogenetic analysis. The results indicated that these strains are closely related to JEC viruses, but forming a distinct subclade together with ILHV, in accordance with results recently reported by Medeiros et al. (2007).

Prevalence of intestinal parasites in a population in south of Tehran, Iran

The aim of this study was to describe epidemiologic characteristics of intestinal parasites in a population in south of Tehran, Iran. A retrospective cross-sectional study of patients with suspicious intestinal parasitic infections referred to the Zakaria Razi Laboratory in Shahre-Ray, southern Tehran, Iran, was conducted from April 21, 2004 to October 20, 2005. All stool samples were examined and socio-demographic informations were retrieved. Of 4,371 referred patients, 466 (239 males and 227 females) were laboratory diagnosed with intestinal parasites, with a period prevalence of 10.7%. Blastocystis hominis (B. hominis) and Giardia lamblia (G. lamblia) were the most frequent intestinal parasites. More than half of patients aged ³ 18 years had a low level of educational attainment (e.g. illiterate, primary school, high school) (170/331, 54.1%). Further, majority of patients were homemakers (42.3%, 140/331) or workers (28.1%, 93/331) employed in various business settings such as food industry and construction. Findings of this study showed that intestinal parasitic infections are still a major public health challenge in Iran that needs to be addressed. We believe that public education, improving sanitation conditions of underdeveloped areas/communities, community involvement, and supporting evidence-based practice/programs are the major keys to success in preventing the spread of intestinal parasitic infections in Iran.

A real-time quantitative assay for hepatitis B DNA virus (HBV) developed to detect all HBV genotypes

Hepatitis B virus (HBV) is a major cause of chronic liver disease worldwide. Besides genotype, quantitative analysis of HBV infection is extensively used for monitoring disease progression and treatment. Affordable viral load monitoring is desirable in resource-limited settings and it has been already shown to be useful in developing countries for other viruses such as Hepatitis C virus (HCV) and HIV. In this paper, we describe the validation of a real-time PCR assay for HBV DNA quantification with TaqMan chemistry and MGB probes. Primers and probes were designed using an alignment of sequences from all HBV genotypes in order to equally amplify all of them. The assay is internally controlled and was standardized with an international HBV panel. Its efficacy was evaluated comparing the results with two other methods: Versant HBV DNA Assay 3.0 (bDNA, Siemens, NY, USA) and another real-time PCR from a reference laboratory. Intra-assay and inter-assay reproducibilities were determined and the mean of CV values obtained were 0.12 and 0.09, respectively. The assay was validated with a broad dynamic range and is efficient for amplifying all HBV genotypes, providing a good option to quantify HBV DNA as a routine procedure, with a cheap and reliable protocol.

Intestinal parasitism and socio-environmental factors among Mbyá-Guarani indians, Porto Alegre, Rio Grande do Sul, Brazil

Disturbing data reveal the prevalence of intestinal parasites and their relationship with socio-environmental factors among Mbyá-Guarani Indians. The prevalence was determined by spontaneous sedimentation in water, centrifugation-floatation, and Kato-Katz. A socioeconomic questionnaire was submitted to each family. The overall prevalence of intestinal parasites was 88.7%, and 45.5% were polyparasitized. There was 90.5% prevalence of enteric parasites in children (1-12- year-old), and 85% among 13-65-year-old individuals, indicating that both age groups are extensively parasitized. The parasite load was low to moderate for geohelminths and 75% of the families did not have latrine, thus the practice of defecation occurred outdoors. These findings suggest that the multiple intestinal parasitism in the Mbyá-Guarani community is high to the point of being the rule, and that it relates essentially to the traditional lifestyle and health habits. It is urgently necessary to implement the association of anti-parasitic treatment with sanitation improvement. This should be done simultaneously with health education activities for this population.

Prevalence and risk factors for intestinal protozoa infection in elderly residents at Long Term Residency Institutions in Southeastern Brazil

This study determined the prevalence of intestinal protozoa in Long Term Residency Institutions for the Elderly (ILPI) in elders, nurses and food handlers, identifying the risk factors associated with the infections. Stool samples taken from the elderly (n = 293), nurses (63) and food handlers (19) were studied. Questionnaires were used with questions related to sociodemographic variables, health, behavior and health characteristics. Stool samples were examined using the techniques of Faust and Ziehl Neelsen, and the prevalence of G. duodenalis, Cryptosporidium spp., E. histolytica/dispar in the elderly was 4.0%, 1.0% and 0.3% respectively. Nurses and food handlers showed 4.8% and 5.2% positivity only for G. duodenalis, respectively. The origin of the individuals and contact with domestic animals has been associated with infection by G. duodenalis in the elderly, and contact with domestic animals was considered a risk factor for infection. The last stool examinations were related to Cryptosporidium spp.. None of the variables were associated with E. histolytica/dispar. The frequency of hand washing was significantly associated with G. duodenalis among nurses. The frequency of positive samples of G. duodenalis, Cryptosporidium spp., E. histolytica/dispar showed that ILPIs environments are conducive to this occurring due to contact between the elderly, nurses and food handlers, which are often poorly trained in hygiene procedures and food handling.

PREVALENCE OF INTESTINAL PARASITISM AND ASSOCIATED SYMPTOMATOLOGY AMONG HEMODIALYSIS PATIENTS

Intestinal parasites are an important cause of morbidity and mortality. Immunocompromised individuals may develop more severe forms of these infections. Taking into account the immunity impairment in patients suffering from chronic renal failure (CRF), we will determine the prevalence and associated symptoms of intestinal parasites in these patients. Controls without CRF were used for comparison. Stool samples were collected and processed for microscopic identification of parasites using the Formalin-ether concentration method. For Cryptosporidium diagnosis, the ELISA technique was used. One hundred and ten fecal samples from hemodialysis patients were analyzed, as well as 86 from a community group used as control group. A result of 51.6% of intestinal parasites was observed in hemodialysis patients and 61.6% in the control group. Cryptosporidium and Blastocystis were the most common infections in patients with CRF (26.4% and 24.5%, respectively). Blastocystis was the most common infection in the control group (41.9%), however no individual was found positive for Cryptosporidium. Among the CRF patients, 73.6% were symptomatic, 54.3% of these tested positive for at least one parasite, in contrast to 44.8% in asymptomatic patients (p = 0.38). The most common symptoms in this group were flatulence (36.4%), asthenia (30.0%) and weight loss (30.0%). In the control group, 91.9% were symptomatic, 60.8% of these tested positive for at least one parasite, in contrast to 71.4% in asymptomatic patients (p = 0.703). A significant difference between the two groups was observed with regard to symptoms, with bloating, postprandial fullness, and abdominal pain being more frequent in the control group than in the hemodialysis group (all p < 0.05). Comparing symptomatic with asymptomatic, there was no association in either group between symptoms or the prevalence of parasitic infection, nor with the type of parasite or with multiple parasitic infections. Patients with chronic renal failure are frequent targets for renal transplantation, which as well as the inherent immunological impairment of the disease itself, results in immunosuppression by medication. For this reason, carriers of intestinal parasites with pathogenic potential can develop serious clinical complications influencing the success of transplantation. This fact, coupled with the high prevalence of intestinal parasites and the dissociation between symptoms and infection in CRF patients, suggests that the stool test should be incorporated in routine propedeutics. Furthermore, preventive measures for the acquisition of parasites through the fecal-oral contamination route should be introduced.

Intestinal parasites in a quilombola community of the Northern State of Espírito Santo, Brazil

The aim of this study was to determine the occurrence of intestinal parasites in a quilombola community from the northern Espírito Santo, Brazil. Descendants of slaves who arrived in Brazil in the sixteenth century, this population settled in the municipality of São Mateus in 1858. Fresh fecal samples from 82 individuals who agreed to participate in the study were collected between August 2009 and July 2010, and immediately sent to the Clinical Laboratory of the Centro Universitário Norte do Espírito Santo of the Universidade Federal do Espírito Santo for analysis. Out of all the participants, 36 (43.9%) were male and 46 (56.1%) were female, whose ages ranged from six to 85 years. The study of the occurrence of intestinal parasites indicated that 35 individuals (42.7%) were infected with at least one intestinal parasite. Among helminths, the most frequent were hookworms, with a rate of 14.6%. With regard to protozoa, Entamoeba coli, Entamoeba histolytica/Entamoeba dispar and Endolimax nana stood out, with frequencies of 23.2%, 8.5% and 4.9%, respectively. The occurrence of biparasitism was observed in 13 of the 82 subjects, accounting for 15.8%, and no cases of multiple parasitic infections were observed. It was concluded that the reduction of cases of intestinal diseases due to parasites will only be achieved with the improvement of basic sanitation and quality of life of quilombola populations.