Indirect Hemaglutination Research Articles

Study of diagnostic value of the reaction of indirect hemaglutination in comparison with enzyme immune assay and other serological tests at cattle brucellosis

The results of comparative brucellosis diagnostic studies of blood sera from cattle at farms with different epizootic situations, performed in 2021 in the laboratory of the Caspian Zonal Research Veterinary Institute – a branch of the Dagestan Agriculture Science Centre, are presented in the paper. The diagnostic efficiency of the indirect hemagglutination test (IHA) was studied in comparison with enzyme immunoassay (ELISA), agglutination test (RA), complement fixation test (CFR), rose-bengal test (RBP) and immunodiffusion reaction with O-PS antigen (RID with O-PS antigen). The specificity of these serological tests was confirmed by the negative results of studies of 40 samples of blood serum of non-vaccinated against brucellosis animals at a brucellosis-safe farm. The sensitivity of the tested diagnostic tools was determined using 46 blood serum samples from cows immunized with the B. abortus 82 vaccine at a brucellosis-prone farm. According to the results obtained, in particular, according to a significant percentage of animals positively reacting to brucellosis in RID with O-PS antigen and high titers of RNHA, RA and RSK, it can be stated that the infection in the herd proceeds in an acute form. Thus, positive results were obtained in RNGA in the study of 18 (39.1%), ELISA (LLC NPF “Sibbiotest”) – 29 (63 %), in RSK – 18 (39.1 %), in RA – 15 (32.6 %), RBP – 15 (32.6 %) and RID with O-PS antigen – 7 (15.2 %) blood serum samples. The conducted studies have shown that RNHA is one of the most sensitive tests, which makes it possible to detect a high percentage of animals with brucellosis at a brucellosis-prone farm (39.1% of those studied) in the early stages after infection and surpasses the results of most other serological methods. The results of comparative serological studies of blood serum samples of cattle in RNHA, IEA, RSK, RA, RBP and RID with O-PS confirmed the specificity of the tests and the high sensitivity of RNHA with brucellosis erythrocyte antigen.

Retrospective Determination of the Prevelense of Anti-<i>Echinococcus granulosus</i> Antibodies in Cystic Echinococcosis Pre-diagnosed Patients at Erciyes University Faculty of Medicine

Cystic echinococcosis (CE) is a parasitic disease that has been known for years in helminth diseases and it is important as human and animal health problem in many parts of the world and in our country due to economic losses. In this study, it was aimed to retrospectively evaluate the distribution of anti-E. granulosus-IgG antibodies in patients with pre-diagnosis of CE that referred to parasitology laboratory between January 2013-December 2018. Commercial kit was used for indirect hemaglutination (IHA), indirect fluorescent antibody test (IFAT) and Western blot (WB) methods using sera from patient samples was applied according to the kit proposal. In addition, patient materials for CAM, CSF and blood for which polymerase chain reaction (PCR)/QPCR tests were requested were examined. Sera of the patients who were tested with at least one of the IHA, IFAT and WB methods or a combination of these methods, and 443 cases out of 2.283 cases were found to be E. granulosus seropositive. It was determined that 369 (62.03%) of 443 positive patients were female and 330 (37.97%) were male patients. Among these patients, 87 patients whose IFAT and/or IHA tests were negative were found to have positive results with the WB method. IFAT or IHA test results of 13 patients with negative WB tests were found to be positive. Four patients were identified with both tests positive but WB test results negative. In addition, 36 of 72 patients who underwent PCR/QPCR tests were found to be positive. As a result of a six-year retrospective screening, 22% of the cases were found to be positive, and it was concluded that the prevalence of CE is high and the use of a single test may be insufficient in the diagnosis of CE, therefore, test combinations will increase the sensitivity and reliability in reaching the correct diagnosis.

ОПРЕДЕЛЕНИЕ БЕЗОПАСНОСТИ ОБЛУЧЕННОЙ ПРОДУКЦИИ ПО СОДЕРЖАНИЮ ХИНОНОВ

The result of research, conducted using our immuno-chemical testing system (Reaction of Indirect Hemaglutination) shows that application of radiation exposure to farming and agro products in dosages needed for prolongation of their storage lifetime, for prevention of rotting and germination or for decontamination of feed-stuff from natural microflora and insects; leads to increase in radiotoxins contents thereof with maximums reached on 7-15 days. Besides, traces of radiotoxins in reaction of indirect hemaglutination depend on type of product, radiation dosage and time of product storage after radiation treatment.

Evaluation of the seropositivity of patients with cystic echinococcosis in Konya, Turkey

Aim: Diagnosis of echinococcosis is based on clinical symptoms, imaging techniques and particularly on the detection of specific antibodies in serum specimens of patients. The aim of this study is retrospective evaluation of seropositivity from patients who is diagnosed as Cystic echinococcosis. Materials and Methods: In this study, the blood serum samples of patients, which were sent to the Medical Microbiology Laboratory from several clinics of the Selcuk University Medical Faculty Hospital between 01 January 2015- 01 July 2020, were evaluated in terms of Cystic echinococcosis by using indirect hemaglutination test. The results were evaluated with a cut off titer ≥160 of as recommended by the manufacturer. Results were evaluated by gender and ages. Results: Cystic echinococcosis (CE) was determined to be seropositive 332 (21.6%) of 1543 patient samples. Seropositivity was found in 226 (14,7%) of 933 female patients and in 106 (6,9%) of 610 male patients. Seropositivity was mostly seen in 21-40 age group. Conclusion: While there have been a number of animal studies, little information is available about the prevalence of Cystic echinococcosis in humans in Turkey. Cystic echinococcosis continues to be a majör public health problem in the Konya province. Large scale prevention and control programs should be implemented against this disease.

Active Transmission of Trypanosoma cruzi in Schoolchildren from the Amazon Region in Napo Province, Ecuador.

Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, is a vector-borne disease with a major disease burden in the Americas, with over 6 million cases. There are about 200,000 cases in Ecuador, but the epidemiology of the disease is poorly understood, particularly in the Amazon region, making surveillance and control challenging. We determined here the seroprevalence of T. cruzi antibodies in a cohort of 516 schoolchildren aged 5-15years from Chontapunta parish, in the Napo province, Ecuador, using ELISA and indirect hemaglutination tests. We detected a seroprevalence of 0.77% (95% confidence interval 0.31-1.97%), with some significant variation among the three studied communities. These data provide evidence of the ongoing transmission of T. cruzi in this area, and support the need to strengthen epidemiological surveillance and patient care.

Evaluation of Usefulness of Three Serological Tests Using Native Crude Antigen in Diagnosis of Hepatic Cystic Echinococcosis Patients

Objective: To evaluate three different serological tests [Indirect Hemaglutination (IHA), Enzyme Linked Immunosorbent Assay (ELISA) and Western Blotting (WB)] using native crude antigen for diagnosis of hepatic cystic echinococcosis (HCE) patients. Materials and Methods: Sheep hydatid fluid (HF) was collected from fertile cysts obtained from a slaughterhouse and used as an antigen. Forty patients who were attended the Dr. Ersin Arslan Training and Research Hospital in Gaziantep, Turkey, were investigated. Serum samples were obtained from surgically confirmed CE patients. Healthy Turkish people and 16 patients with other helminthic infections were included as a control group. Results: Of the 40 analyzed patients, 10 (25%) were men and 30 (75%) were female. The average age was 46.97 years (s.d.; 18.95). The majority of the patients had a single cystic lesion situated in one lobe of the liver (usually in the right lobe) (55%), 32.5% of patients had two cystic lesions and 12.5% of patients had multiple cyst formations with various numbers. In all cases, ultrasound (US) examinations were positive and the size of cysts was between 2.1 - 12.7 cm. Twenty-three patients of the total 40 patients were classified according to the WHO classification system based on US findings. According to the results of WB analysis, molecular weights of 8 kDa (80%), 12 kDa (80%), 22 - 24 kDa (97.5%), 26 kDa (97.5%), 34 kDa (100%), 36 - 38 kDa (90%), 45 - 50 - 55 kDa (97.5%), and 60 - 75 kDa (97.5%) bands were identified. But 34, 50, and 55 kDa bands were also found in other helminthic diseases. Conclusion: The specificity and sensitivity of three serological tests (IHA, ELISA and WB) using crude antigen were compared by diagnosing hepatic cystic echinococcosis patients. IHA and ELISA showed high sensitivity but low specificity. Western blotting showed low sensitivity but high specificity.

Surgical management of hepatic hydatid cyst in children

The aim of this study was to demonstrate the surgical treatment and outcome of hydatid cyst in liver in children. The age of presentation varied from 8-11 years. All patients had abdominal pain, five had lump in right hypochondrium and three patients had fever. Hydatid cyst serology, indirect hemaglutination test, ultrasonography and CT scan done in all the patients. Histological examination supported the diagnosis of hydatidosis since the cyst wall contains outer laminated and inner germinal layer. All the patients got relief of their preoperative symptoms after surgery. Two patients had positive serology after surgery. Therefore, they were given two additional regimen of albendazole and turned out to be serologically negative. In conclusion, hydatid cyst management in children is not consensual. However, conservative surgery remains the treatment of choice to avoid serious complication in the endemic region of Indian subcontinent.

Http://www.pubvet.com.br/artigo/2888/p-styletext-align-justify-aligncenterstrongavaliaccedilatildeo-laboratorial-da-sauacutede-de-macacos-prego-emcebus-apellaem-na-cidade-de-jataiacute-gostrongp

The aim with this study was to evaluate, with laboratory tests, health of five capuchin monkeys living in a public park on the Diacuí Lake in the city of Jataí-GO, Brazil. The mean values for the main hematological parameters were 5.82 x 106/uL; 41.05%; 77.50 x 103/uL; and 7800/uL, respectively for red blood cells; hematocrit; platelets; and white blood cells count. It was not found any hematozoa in blood smears. In biochemical parameters, average concentrations of urea; creatinine; alanine aminotransferase; glucose; and total cholesterol were, respectively, 25.68 mg/dL; 0.79 mg/dL; 55.00 U/L; 96.16 mg/dL; and 87.16 mg/dL. None of the animals showed up reagent to the indirect hemaglutination test for antibodies to Toxoplasma gondii. Thus, we can conclude that these animals were seen to be in good health at the time of collection of blood samples.

PRESENCIA DE INFECCIÓN POR Trypanosoma cruzi Y Toxoplasma gondii EN PERROS DOMÉSTICOS DE LOCALIDADES RURALES EN EL NOROESTE ARGENTINO

Zoonoses caused by the protozoan parasites Trypanosoma cruzi and Toxoplasma gondii are prevalent in Argentina, where dogs are reservoir hosts but also suffer Chagas and Toxoplasmosis disease. Since the dynamic of both diseases in human populations is reflected in canines, knowing the seroprevalence of these zoonoses in dogs is considered a good strategy to understand disease patterns. We aimed to know the seroprevalence of T. gondii and T. cruzi infection in canines from four rural areas in the Salta province by Indirect Hemaglutination (IHA) and Enzyme-Linked Immuno Sorbent Assay (ELISA). Serologic results from 209 dogs showed 18.2% (4/22) presence of anti- T. gondii antibodies in the locality of La Union, department of Rivadavia, 20.5% (8/39) in San Carlos, 13.1 % (11/84) in Cafayate, and 23.4% (15/64) in Embarcacion department of General San Martin. Anti- T cruzi antibodies were only detected in San Carlos, being 28.2% (11/39). Frequent seroprevalence to T. gondii , found in dogs of all locations sampled suggest environmental pollution by this parasite in the area. The discovery of infected dogs by T. cruzi , in the town of San Carlos, suggests the possibility of active transmission of the parasite in the recent past and alerts us to the possibility of reservoir hosts.

Diagnostic parameters of serological ELISA for invasive amoebiasis, using antigens preserved without enzymatic inhibitors

Amoebiasis is the third cause of death due to parasites in the world. Although, numerous serodiagnostic and salivary tests have been developed, the majority of these assays lack sensitivity in endemic zones to detect acute amoebic liver abscess. The two main limiting factors to develop reliable assays are the high levels of anti-amoeba antibodies in populations living in endemic zones, and the proteolysis of amoebic extracts even treated with inhibitors. Our group reported a method to preserve amoebic antigens without using enzymatic inhibitors (IC:MC fraction) that shows stability for years. Here we describe the development of a serologic ELISA to diagnose amoebiasis made with IC: MC antigens, and its validation for clinical use in endemic areas. In our study, we included sera from 66 patients diagnosed with acute amoebic liver abscess and 33 volunteers living in an endemic area for amoebiasis. Our assay was compared with an indirect haemaglutination assay (IHA) an ELISA elaborated with antigens derived from untreated trophozoites. The ELISA made with IC: MC antigens presented more reproducibility compared to other assays. Sera from 95% ALA patients showed a positive value. The ELISA (IC: MC) detected 97% of patients with ALA compared to an 81% using IHA. The parameters of ELISA (vs. IHA) were Sensitivity 98% (81%), Specificity 96% (97%), Positive predictive value 98% (96%), Negative predictive value 96% (73%) and Accuracy 98% (87%). A negative serologic test does not rule out the diagnosis of invasive amoebiasis. The ELISA made with antigens preserved without using enzymatic inhibitors has valuable serodiagnostic value to diagnose acute amoebic liver abscess, even in populations living in endemic zones of amoebiasis carrying antibodies against amoebas. In conclusion, ELISA-IC:MC presented better diagnostic parameters than IHA although a negative serologic test does not rule out acute invasive amoebiasis.

Seroprevalence of Trypanosoma cruzi Infection in Schoolchildren and in Pregnant Women from an Amazonian Region in Orellana Province, Ecuador.

Chagas disease is a parasitic disease caused by the protozoan parasite Trypanosoma cruzi and about 230,000 persons are estimated to be infected in Ecuador. However, limited studies have been performed in the Amazon region, on the eastern side of the country. We evaluated here the seroprevalence of Trypanosoma cruzi infection in 12 rural villages of the Loreto canton, Orellana Province in schoolchildren aged 5-15 years and in pregnant women. A total of 1,649 blood samples were tested for Trypanosoma cruzi antibodies by enzyme-linked immunosorbent assay and indirect hemaglutination, and discordant samples were tested by indirect immunofluorescence assay. We detected a seroprevalence of anti-Trypanosoma cruzi antibodies of 1.3% in schoolchildren aged 5-15 years, indicating the persistence of a constant and active vectorial transmission in the Loreto County and confirming the need of the implementation of nonconventional vector control. We also observed a seroprevalence of 3.8% in pregnant women, indicating a clear risk of congenital transmission. Further studies should help define this risk more precisely and implement current international guidelines for the diagnosis, treatment, and care of these cases.

Evaluation of Cystic Echinococcosis Suspected Patients Applied to National Parasitology Reference Laboratory of Public Health Institution of Turkey Between 2009-2013

Cystic echinococcosis (CE) caused by the metacestode form of Echinococcus granulosus is an important public health problem common in our country. In this study, anti-E. granulosus antibodies were aimed to investigate in the serum samples of CE suspected patients who applied to the National Parasitology Reference Laboratory of Public Health Institution of Turkey. In the study, serum samples of 2921 patients which were sent to our laboratories from different hospitals between 1 January 2009 and 31 December 2013 were evaluated with at least one of the following tests: Enzyme-Linked Immunosorbent Assay (ELISA), Indirect Hemaglutination Assay (IHA) and Western Blot (WB) techniques. Four hundred thirty nine (15.03%) of inspected 2921 samples were determined seropositive with at least one of the methods. When the results analyzed by gender, 13% of males and 16.40% of females were found positive. Examined the distribution of the results by years, with a maximum of 25% positivity was observed in 2009. Compatibility was determined at the rate of 91.4% among ELISA and IHA results; also 89.7% among WB and the other tests results. Despite the gradual decreases the CE in Ankara and its surroundings, it is still continues to be a major public health problem. Essential prevention and control measures should be taken to reduce the prevalence of the disease. Also in the diagnosis of the disease, more reliable results can obtained with applying two tests (ELISA/IHA) together and confirm the positivity with WB.

Evaluation of the Chagas disease control program in Açucena Municipality, Rio Doce Valley, State of Minas Gerais, Brazil.

Açucena Municipality, Rio Doce Valley, State of Minas Gerais, Brazil temporarily (2001-2005) interrupted epidemiological surveillance for Chagas disease. The objective of this work was to evaluate the Chagas Disease Control Program (CDCP) in Açucena and to offer suggestions for improving local epidemiological surveillance. This study was conducted in three phases: I) a serological investigation of schoolchildren aged 5 to 15 years using an enzyme-linked immunosorbent assay (ELISA) test performed on blood collected on filter paper followed by ELISA, indirect immunofluorescence (IIF) and indirect hemaglutination (IHA) on venous blood for borderline cases and those in the gray zone of reactivity; II) vector evaluation using the data obtained by local health agents during 2006-2010; and III) examination by ELISA, IIF and IHA of serum samples from the inhabitants of houses where infected Triatoma vitticeps was found and evaluation of their knowledge about Chagas disease. Five individuals had inconclusive results in the ELISA screening but were seronegative for Chagas disease. The triatomine evaluation revealed the presence of three species: Triatoma vitticeps, Panstrongylus megistus and Panstrongylus diasi. Triatoma vitticeps was the most prevalent and widespread, with a higher (67%) index of Trypanosoma cruzi flagellates and evidence of colonization. Most of the inhabitants of the infested houses recognized triatomines and had basic knowledge about Chagas disease. Although T. vitticeps is not clearly associated with Chagas disease transmission, these results highlight the importance of maintaining CDCP in endemic areas and the need for greater emphasis on epidemiological surveillance, especially in areas with important vectorial changes or that have been modified by human intervention.

DETERMINACIÓN DE ANTICUERPOS CONTRA Toxoplasma gondii EN BÚFALOS DE AGUA (Bubalus bubalis) EN EL DISTRITO DE JENARO HERRERA, LORETO, PERÚ

The aim of this study was to determine the seroprevalence of Toxoplasma gondii in buffalos in the Jenaro Herrera district, Loreto, Peru. A total of 70 blood samples were collected in female buffalos for the detection of antibodies against T. gondii by the Indirect Hemaglutination Test (IHA) and the Indirect Immunofluorescence Test (IFI). The results showed that 35.7 ± 11.2% of the samples were positive by IHA and 17.1 ± 8.8% were positive by IFI.

Tuboovarian Abscess Caused by Hydatid Cyst: A Rare Case

Primary lesions of hydatid cysts caused by Echinococcus granulosus, are frequently localized in liver, followed by lungs, muscles, kidneys, spleen and bones. Pelvic inoculations are rare and usually occur as a secondary infection. In this report, a case of primary hydatid cyst in the abdomen, spleen and pelvic organs, clinically mimicking tuboovarian abscess, was presented. A nineteen-years-old female patient was admitted to the gynecology outpatient clinic with the complaint of abdominal pain for two days. The case was considered as tuboovarian abscess according to the initial examination findings and hospitalized for treatment and follow-up. In transabdominal ultrasound examination, 44 x 43 mm thin-walled septated cysts in the left ovary and 65 x 65 mm thin-walled multiloculated cysts in the spleen were detected. Abdominal computerized tomography also yielded multivesicular cystic masses in spleen, front abdominal wall and the left ovary. Since the clinical and vital findings worsened, she initially underwent ovarian cystectomy by laparoscopy, then abdominal cystectomy and splenectomy. The operation material examined macroscopically was compatible with hydatid cyst with the characteristics of a germinative membrane and hydatid sand. The diagnosis was confirmed by histopathological examination. The patient was discharged without complication on post-operative sixth day, with a recommendation of albendezol (15 mg/kg/day, 3 months) treatment. Since the patient had undergone emergency surgery, indirect hemaglutination (IHA) test had not been performed pre-operatively. However, post-operative third month serum sample revealed a positive (1/32) IHA titer. In conclusion, hydatid cyst should be kept in mind in the differential diagnosis of patients with abdominal pain, in response to the high prevalence of the parasite in our country.

DETERMINACIÓN DE LA SEROPREVALENCIA DE TOXOPLASMOSIS EN ALPACAS Y LLAMAS EN LA ESTACIÓN EXPERIMENTAL INIA-PUNO

The objective of this study was to determine the seroprevalence of Toxoplasma gondii in alpacas and lamas from the experimental station of INIA-Puno (Quimsachata), located in the Lampa province, Puno. For this purpose, 200 alpacas and 136 lamas, both male and female, were sampled in June 2000. Blood serum was collected to detect antibodies by the indirect haemaglutination test (IHA). 44.5 ± 6.9% (89/200) of alpaca samples had antibodies. Seroprevalence found in females (56.0 ± 9.7%) was higher than that in males (33.0 ± 9.2%) (p<0.01). Seroprevalence increased with age (p<0.01). The seroprevalence in lamas was 27.9 + 7.5% without differences between sexes, but increasing with age (p<0.01). The results showed a seroprevalence relatively higher in alpacas than in lamas at the INIA-Puno station. It was found that sex is a risk factor for acquiring toxoplasmosis in alpacas, but not in lamas, and age is a risk factor for the infection with T. gondii for both alpacas and lamas (p<0.01).

FRECUENCIA DE ANTICUERPOS CONTRA Toxoplasma gondii EN MONOS CEBUS APELLA CRIADOS EN CAUTIVERIO.

The purpose of the present study was to determine the frequency of primates (Cebus apella) carrying anti-Toxoplasma gondii antibodies at the zoo of the Patronato del Parque de las Leyendas, in Lima. Sixty two sera samples were obtained and analyzed by the Indirect Hemaglutination test (HAI) to detect antibodies against T. gondii. The results showed that 90.3% (56/62) sera were positive, and no significant asociation due to sex and age (juvenile and adult) were found. The high rate of positive primates to this parasite could be due to the high number of domestics cats in the premises.

FRECUENCIA DE Toxoplasma gondii EN GATOS DE LIMA METROPOLITANA Y CONCORDANCIA ENTRE LAS TÉCNICAS DE INMUNOFLUORESCENCIA INDIRECTA Y HEMAGLUTINACIÓN INDIRECTA

The aim of the present work was to estimate the frequency of antibodies anti- Toxoplasma gondii in cats of Lima city, and to determine the rate of concordance between the Indirect Immunofluorescence (IFI) and Indirect Haemaglutination (HAI) assays. A total of 178 cat serum samples were collected in veterinary clinics of varios districts of Lima. The results showed 11.2 ± 4.6% (HAI) and 17.9 ± 5.6% (IFI) positive serums to T. gondii, and without statiscal differences due to sex and age group. The Kappa (K) value was 0.73 indicating a substantial concordance between the two assays indicating that both assays can be reciprocally replaced.

CONCORDANCIA ENTRE LAS PRUEBAS DE HEMAGLUTINACIÓN INDIRECTA E INMUNOFLUORESCENCIA INDIRECTA PARA DETERMINAR LA PREVALENCIA DE Toxoplasma gondii EN OVINOS.

The objective of this study was to measure the degree of concordance between the Indirect hemaglutination (HAI) and Indirect immunofluorescence (IFI) tests for Toxoplasma gondii detection in sheep. Atotal of 310 sera were collected from ewes of different ages at Fundo SantaMaría, district ofNuñoa, province ofMelgar, Puno. TheKappa (K) value was 24%, showing that the degree of association between both techniques was of a regular type. TheMcNemar test showed significative differences between both tests, so one cannot replace the other. The seroprevalence for T. gondii was 50.3 ± 5.6% through HAI test and 88.1 ± 3.5%through IFI test. Seroprevalence in animals &lt;1, 1-2, 2-3, and &gt;3 years of age was 23.2, 33.3, 52.1, and 58.7% through HAI test and 72.0, 90.0, 91.2, and 90.5% respectively through IFI test.

SEROPREVALENCIA DE Toxoplasma gondii EN ALPACAS (Lama Pacos) EN UNA UNIDAD DE PRODUCCIÓN DE LA SIERRA CENTRAL DEL PERÚ.

The aim of this study was to determine the seroprevalence of Toxoplasma gondii in alpacas from a commercial unit of SAIS Tupac Amaru, located at Junín Department. A total of 200 blood samples were collected in alpacas of both sexes for the detection of antibodies against T. gondii by the Indirect Hemaglutination Test (IHA). The results indicated that 21 ± 5.6% (42/200) of the samples had antibodies and the titres varied from 1/16 till 1/1024. The Logistic Regression Test failed to obtain statistical relationship between the presence of T. gondii antibodies and the sex of the animal, but there was a significant relationship with age. Young alpacas (8 to 12 months of age) were 2.9 times more susceptible than the groups of &gt;12-36 and &gt;36 months of age. These results could be due to the presence of maternal antibodies, animal management, environmental factors, and the presence of the definitive host. The prevalence found in the present study was considered as moderate.