Detection of IgG Anti-Giardia duodenalis Antibodies in Sera by Indirect Immunofluorescence and Western Blotting Assays.

INTRODUCTION: Serological assays are alternative laboratory tools for the diagnosis of parasitic infections. The aim of this work was to evaluate the performance of the indirect fluorescent antibody test (IFAT) and Western blotting (WB) for the detection of IgG anti-Giardia antibodies in human sera. METHODOLOGY: Sera from individuals infected with Giardia duodenalis, other parasites or non-parasitized were selected for serological assays. Ninety-seven sera were tested by IFAT at 1:20 and 1:40 dilutions and of these, 40 samples were also analyzed by WB. RESULTS: The sensitivity and specificity of the IFAT was 97% and 46.9% at 1:20 sera dilution, and 39.4% and 59.4% at 1:40 sera dilution. The low molecular weight polypeptides fractions of 25 kDa, 27-31 kDa and 45-55 kDa were the most frequently identified by the sera of individuals infected with G. duodenalis, along with low cross-reactivity, presenting an individual sensitivity of 42.8%, 50.0% and 57.1%, and specificity of 83.3%, 83.3% and 91.7%, respectively. The highest overall sensitivity of WB (85.7%) was based on the immunoreactivity of sera with at least one of those proteins. The concordance between the detection of G. duodenalis in feces by microscopy and the WB results was considered substantial (Kappa = 0.61). CONCLUSION: Constant exposure to Giardia infection throughout a lifetime can maintain high levels of specific antibodies in serum, even without active infection. Moreover, proteins found in intestinal amoebas may hinder the serological diagnosis of giardiasis in endemic areas due to cross-reactivity, which can be partially solved using Giardia low molecular weight proteins.

Seroepidemiological Survey of Chronic Chagas Disease in a Rural Community in Southern Bahia, Brazil, Using Recombinant Chimeric Antigens.

Chagas disease (CD), caused by the parasite Trypanosoma cruzi, is a neglected tropical disease with life-threatening implications. In this study, we conducted a seroepidemiological survey to determine the prevalence and clinical profiles of CD in 217 individuals from an impoverished rural community in Southern Bahia, Brazil. The overall prevalence of CD in the studied community was 0.92%, detected through latent class analysis (LCA). Two individuals tested positive for anti-T. cruzi IgG, both being male farmers. One case was a 22-year-old man born in Camamu, with no evidence of congenital transmission, suggesting other routes of transmission such as vector-borne transmission due to migratory activities. The other case was a 69-year-old man born in São Felipe, who had lived in an adobe/brick house and had a pacemaker due to cardiac involvement caused by CD. The prevalence in this community was lower than expected, given the socioeconomic conditions and environmental factors that contribute to T. cruzi transmission. This could be attributed to the implementation of preventive measures and vector control programs by the Brazilian Government. However, continuous monitoring and surveillance are essential to sustain control efforts and detect any potential re-emergence of the disease. While the overall prevalence was low, the detection of positive cases underscores the need for continued surveillance and control measures in vulnerable populations, such as rural communities. Active surveillance, early diagnosis, and timely treatment are crucial in preventing disease progression and complications, thereby enhancing the effectiveness of screening and treatment programs.

Modulation of circulating cytokine production in alcoholic patients infected with Strongyloides stercoralis.

Strongyloidiasis control is associated with a Th2 immune response. However, alcohol ingestion plays an important role in modulating the immune system. The aim of this study is to evaluate the occurrence of Strongyloides stercoralis infection in alcoholic patients, the levels of circulating cytokines (IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-15 and IL-17), and its correlation with modulation of parasitic load in alcoholic individuals infected with S. stercoralis. A total of 336 alcoholic patients, treated at the Alcoholic Care and Treatment Center were included in this study. The cytokine levels were measured by a commercial ELISA in 80 sera divided into four groups with 20 individuals each: alcoholics infected (ASs+) and not infected (ASs-) with S. stercoralis and non-alcoholics infected (NASs+) and not infected (NASs-) with the helminth. S. stercoralis frequency in alcoholic patients was 16.1% (54/336). The parasitic load varied from 1 to 546 larvae/g of faeces, median and interquartile range (IQR) of 9 and 1.0-62.5 larvae/g of faeces, while in non-alcoholic individuals the parasitic load was less than 10 larvae/g of faeces. Levels of circulating IL-4 were significantly higher in ASs+ when compared with NASs- group (p < .05). An inverse correlation between serum levels of IFN-γ and parasitic load in alcoholic patients infected with S. stercoralis was observed (r = -601; p < 0.01). These results suggest that modulation of IFN-γ production occurs in alcoholic individuals with high parasitic burden.

Ineffectiveness of TF-Test® and Coproplus® Methods in Strongyloides stercoralis Infection Diagnosis.

PURPOSE: S. stercoralis diagnosis is currently performed by parasitological methods, mainly by Baermann-Moraes (BM), although Agar Plate Culture (APC) presents a higher sensitivity. New techniques, such as TF-Test® and Coproplus® have been suggested as more practical alternatives. The aim of this study was to evaluate the sensitivity of TF-Test® and Coproplus®, compared with APC, BM and Spontaneous Sedimentation (SS) methods. METHODS: One-hundred and forty male alcoholic patients, who provided three stools samples collected on alternate days, were included in this study. The most frequently found parasite was S. stercoralis, 20% (28/140), and the most sensitive method was APC, 96.4% (27/28), followed by BM, 89.3% (25/28) and SS, 57.1% (16/28). TF-Test® and Coproplus® presented a sensitivity of 46.4 (13/28) and 39.3% (11/28), respectively. In samples with a parasitic load of 1-10 larvae/g of feces, which occurred in 39.3% (11/28) of the infected patients, both the TF-Test® and Coproplus® methods demonstrated sensitivities of 18.2% (2/11), while APC and BM methods reached a sensitivity of 100% (11/11) (p < 0.05). For other intestinal helminth infections, TF-Test® and Coproplus® sensitivities were 22.2 (4/18) and 11.1% (2/18), respectively, this being lower than the SS, 66.7% (12/18) (p < 0.05). On the other hand, for protozoa infection diagnosis, TF-Test® and Coproplus® presented the highest sensitivities, 62.2 (46/74) and 43.2% (32/74), respectively. CONCLUSION: TF-Test® and Coproplus® methods presented the lowest sensitivities for S. stercoralis and other helminth infection diagnosis; therefore, they can be indicated for use in parasitological diagnosis, only when associated with other more effective methods of helminth identification.

Development of rSs-NIE-1 and rSs-IR Recombinant Antigen-Based Immunoblot for Detection of Antibody to Strongyloides stercoralis.

Strongyloides stercoralis is a soil-transmitted nematode that can cause life-threatening conditions in immunocompromised persons. In the United States, strongyloidiasis should be considered mainly in immigrants, refugees, or travelers. The confirmatory laboratory diagnosis is usually performed by detecting larvae from the stool, duodenal material, and sputum. In persons who are immunocompromised with severe strongyloidiasis, adult worms and eggs can be detected from duodenal material. For serological diagnosis, most assays use crude antigens to detect anti-S. stercoralis IgG. Recently, recombinant proteins such as rSs-NIE-1 and rSs-IR have been used to detect IgG antibodies. We used rSs-NIE-1 and rSs-IR recombinant antigens to develop a biplex Western blot assay to detect the IgG4 antibody in individuals with strongyloidiasis. The sensitivities of rSs-NIE-1 and rSs-IR were 97.4% and 90.8%, respectively, whereas the specificities were 97.6% and 98%, respectively. In conclusion, the biplex rSs-NIE-1 and rSs-IR immunoblot performs well in detecting IgG4 antibody in S. stercoralis-infected persons.

Alcohol consumption alters anti-Strongyloides stercoralis antibodies production.

Individuals infected with Strongyloides stercoralis have been reported to produce different immunoglobulins isotypes, yet few studies have evaluated their use in strongyloidiasis diagnosis. The aim of this work was to evaluate the immunoreactivity of different classes and subclasses of anti-S. stercoralis circulating antibodies in alcoholic patients by ELISA and to perform immunoblotting in samples with discordant results between parasitological and immunological methods. 345 male patients with a clinical diagnosis of alcoholism hospitalized at a reference center for alcoholics in Salvador, Bahia, Brazil, were included in this study. The fecal samples were examined by three different parasitological methods (spontaneous sedimentation, Baermann-Moraes and Agar Plate Culture methods). The ELISA was performed for the detection of IgG, IgG1, IgG4, IgE and IgA1 anti-S. stercoralis. Immunoblotting, for the detection of specific IgA1, was used to elucidate discordant results between parasitological and immunological methods. S. stercoralis infection frequency in alcoholic patients by parasitological methods was 21.4% (74/345). Although IgE-ELISA demonstrated a high sensitivity and specificity in non-alcoholic patients, about 30% (22/74) of alcoholics with larvae in feces were negative. IgG1-ELISA detected the lowest frequency of antibodies in alcoholic patients with larvae in feces, only 57% (42/74). IgG4-ELISA was the best assay for S. stercoralis infection immunodiagnosis. Immunoreactivity in the immunoblotting for IgA1 at 90, 75, 26 and/or 17 kDa bands was observed in 92% (33/36) of alcoholics with larvae excretion and negative ELISA for one or more antibody isotypes. In conclusion, IgG4-ELISA showed the highest sensitivity and specificity, thus demonstrating its superiority for strongyloidiasis immunodiagnosis in alcoholic and non-alcoholic individuals. Both, IgE and IgG1-ELISA presented high sensitivities and specificities for S. stercoralis infection diagnosis in non-alcoholics, however there was low reactivity in alcoholic individuals. This can be associated with an increased susceptibility to severe strongyloidiasis in these patients. IgA1-immunoblotting can be used to confirm S. stercoralis infection when there are discordant results between parasitological methods and ELISA.

Case Report: Strongyloides stercoralis Hyperinfection in a Patient with HTLV-1: An Infection with Filariform and Rhabditiform Larvae, Eggs, and Free-Living Adult Females Output.

Strongyloides stercoralis is the main etiological agent of human strongyloidiasis. Severe strongyloidiasis is commonly associated to alcoholism, corticostereoid use, and human T cell lymphotropic virus type 1 (HTLV-1) coinfection. Herein, we report a case of a 13-year-old boy coinfected with S. stercoralis and HTLV-1, excreting several parasitic forms in the stool. The parasitological examination of his feces showed a large amount of filariform (about 3,000 larvae per gram of feces) and rhabditiform larvae (about 2,000 larvae per gram of feces). In addition, free-living adult females (about 50 parasites per gram of feces) and eggs (about 60 eggs per gram of feces) were detected. The main laboratory findings pointed to high immunoglobulin E (IgE) levels (228 UI/mL) and eosinophila (11.6%). The patient was treated with three courses of ivermectin (200 µg/kg twice, 2 weeks apart), achieving the parasitological cure. An increase of about 19 times in interleucin (IL)-17 level was observed following the parasitological cure, in addition to a decrease in the white blood cell, eosinophil counts, and IgE levels. This is the first case report, to our knowledge, in which an S. stercoralis adult free-living female was described in human feces and where an increase in IL-17 levels after Strongyloides treatment in a HTLV-1 coinfected individual was observed. This finding raises the need for further studies about IL-17 immunomodulation in S. stercoralis and HTLV-1 coinfected patients.

Alterations in serum paraoxonase-1 activity and lipid profile in chronic alcoholic patients infected with Strongyloides stercoralis.

The objective of this study was to investigate paraoxonase-1 (PON1) activity, cortisol levels, and the lipid profile in the sera of alcoholic and non-alcoholic Strongyloides stercoralis-infected and uninfected individuals in a sample of 276 individuals attended at the National Health System in Salvador, Bahia, Brazil. The activity of PON1 was measured by the Beltowski method, serum lipids, and cortisol levels using commercial kits. PON1 activity was low in both alcoholic and non-alcoholic individuals infected with S. stercoralis. A positive correlation was observed between PON1 activity and cortisol concentration in alcoholic individuals who were not infected with S. stercoralis; whereas a negative correlation occurred in S. stercoralis-infected nonalcoholic individuals. The levels of triglycerides, LDL-C, and VLDL-C in S. stercoralis-infected alcoholic individuals were significantly lower than in uninfected alcoholic individuals. The high level of HDL-C and the low level of LDL-C, VLDL, triglycerides and PON1 activity in alcoholic patients infected with S. stercoralis evidenced an anti-atherogenic pattern.

Strongyloides stercoralis infection in patients with systemic lupus erythematosus: diagnosis and prevention of severe strongyloidiasis.

AIM: Strongyloides stercoralis infection is usually chronic and asymptomatic and may persist undiagnosed for decades. However, in immunocompromised individuals, the infection can cause hyperinfection and dissemination. Therefore, early diagnosis is essential to prevent severe forms of strongyloidiasis. The aims of this study were: (i) to evaluate the frequency of S. stercoralis infection in patients with systemic lupus erythematous (SLE) and (ii) to estimate specific immunoglobulins G (IgG) and E (IgE) production using an enzyme-linked immunosorbent assay (ELISA) method. METHODS: Seventy-five SLE patients treated with prophylactic anthelmintic therapy were evaluated using the spontaneous sedimentation (SS), Baermann-Moraes (BM) and agar plate culture (APC) methods. Serum anti-S. stercoralis IgG and IgE antibodies were measured by ELISA. RESULTS: Using parasitological methods, the frequency of intestinal parasites was 10.7%, whereas the frequency of S. stercoralis infection was 1.3%. The sensitivity of the ELISA to detect anti-S. stercoralis IgG and IgE was 80% and 76.9%, respectively. Both assays presented the same specificity of 96.7%. The frequency of anti-S. stercoralis IgG and IgE was 16% and 28%, respectively. Six patients were positive for both antibodies. CONCLUSIONS: Diagnostic approaches using high-sensitivity parasitological methods and the detection of specific antibodies are essential for the diagnosis of strongyloidiasis in immunocompromised patients. Early detection of infection can alter the course of the disease via appropriate treatment, preventing the occurrence of severe strongyloidiasis.

Recurrence of strongyloides stercoralis infection in a patient with Hansen's disease: a case report.

In patients with immunosuppressive disorders, S. stercoralis infection may develop into a hyperinfection syndrome which, on rare occasions, may be a life-threatening condition. Therapy of S. stercoralis infection with thiabendazole has been limited, due to its numerous side effects, and has been replaced by albendazole and ivermectin. The present case report describes a case of Strongyloides Hyperinfection Syndrome (SHS) in a patient with Hansen's disease and lack of response to first-line anthelmintic treatment. A 38 year-old man was diagnosed as having borderline lepromatous leprosy. He developed Erythema Nodosum Leprosum and was treated with thalidomide and prednisone. In May 2010 he was diagnosed with S. stercoralis infection and was treated with albendazole. One year later, the stool examination showed continued presence of S. stercoralis larvae. He was treated with ivermectin (6 mg) in a double dose (given 1 month apart) which resulted in larvae excretion clearance. The absence of infection was confirmed three times during a 1 year followup period by stool examination and non-detection of anti-S. stercoralis IgG levels.