Application of LAg-avidity EIA in the estimate of HIV-1 incidence among key surveillance populations in Hubei Province / 公共卫生与预防医学

Objective Limiting antigen avidity enzyme immunoassay (LAg-Avidity EIA) was applied to calculate the HIV-1 incidence of new HIV infections among men who have sex with men(MSM), drug users(DUS), female sex workers(FSW) and male patients in sexually transmitted diseases clinics(STD) in 2017-2020 of Hubei. Methods HIV-1 antibody positive samples were collected from four key groups in the national HIV sentinel surveillance from 2017 to 2020, and tested by the LAg-Avidity EIA to estimate the annual new infection rate of each group. Results The prevalence of MSM was 2.90%, 4.48%, 3.63 and 2.78% from 2017-2020, respectively, which was the highest among the four groups. The rate of late diagnosis increased from 16.67% in 2017 to 28.81% in 2020.The incidence of MSM was 1.64%(0.85%-2.44%),1.41%(0.59%-2.23%),1.46% (0.71%-2.21%) and 0.96%(0.34%-1.57%), respectively, and the difference was not statistically significant (χ2=1.491, P=0.684). The prevalence of STD from 2017-2020 was 0.03%, 0.01%, 0.06% and 0.08%, respectively, with the incidence was 0.09% (-0.06%-0.25%), 0.10(-0.06%-0.26%), 0.26% (0.02%-0.50%) and 0.04%(-0.07%-0.15%). The prevalence and incidence among FSW remains very low, while the prevalence of DUS fluctuated slightly, but no new infections have been detected. Conclusion The incidence of new HIV infection among MSM in Hubei province from 2017-2020 is relatively high compared with other key groups, but the overall trend is declining, thus the intervention of MSM should be further strengthened and expand the coverage of testing in key populations.

IgG Avidity in Samples Collected on Filter Paper: Importance of The Early Diagnosis of Congenital Toxoplasmosis / Avidez de IgG em amostras coletadas em papel filtro: Importância no diagnóstico precoce da toxoplasmose congênita

Abstract Objective The purpose of the present study is to standardize and evaluate the use of the immunoglobulin G (IgG) antibody avidity test on blood samples from newborns collected on filter paper to perform the heel test aiming at its implementation in ongoing programs. Methods Blood samples from newborns were collected on filter paper simultaneously with the heel prick test. All samples were subjected to immunoglobulin M IgM and IgG enzyme-linked immunosorbent assays (ELISA). Peripheral blood was collected again in the traditional way and on filter paper from newborns with high IgG levels (33). Three types of techniques were performed, the standard for measuring IgG in serum, adapted for filter paper and the technique of IgG avidity in serum and on filter paper. The results of the avidity test were classified according to the Rahbari protocol. Results Among the 177 samples, 17 were collected in duplicate from the same child, 1 of peripheral blood and 1 on filter paper. In this analysis, 1 (5.88%) of the 17 samples collected in duplicate also exhibited low IgG avidity, suggesting congenital infection. In addition, the results obtained from serum and filter paper were in agreement, that is, 16 (94.12%) samples presented high avidity, with 100% agreement between the results obtained from serum and from filter paper. Conclusion The results of the present study indicate that the avidity test may be another valuable method for the diagnosis of congenital toxoplasmosis in newborns.

Resumo Objetivo O objetivo do presente estudo é padronizar e avaliar a utilização do teste de avidez de anticorpos imunoglobulina G (IgG) em amostras de sangue de recémnascidos (RNs) coletadas em papel filtro para a realização do teste do pezinho visando a implementação nos programas já vigentes. Métodos Foram coletadas amostras de sangue de recém-nascidos em papel filtro simultaneamente ao teste do pezinho. Em todas as amostras, foram realizados os testes imunoenzimáticos (ELISA) imunoglobulina M (IgM) e IgG. Dos RNs que apresentaram altos índices de IgG (33), foi novamente coletado sangue periférico da forma tradicional e em papel filtro. Foram realizadas técnicas padrão para a dosagem de IgG em soro, adaptadas para papel filtro, e a técnica de avidez de IgG em soro e em papel filtro. Os valores obtidos para o teste de avidez foram classificados de acordo com o protocolo de Rahbari. Resultados Dentre as 177 recoletas, em 17 amostras foi realizada a coleta simultânea de sangue periférico e papel filtro da mesma criança. Nesta análise, 1 (5,88%) das 17 amostras coletadas em duplicata obteve também baixa avidez de IgG, sugerindo infecção congênita da criança, e houve concordância entre os resultados obtidos em soro e em papel filtro: 16 (94,12%) das amostras apresentaram alta avidez, com concordância de 100% entre os resultados obtidos em soro e em papel filtro. Conclusão Os dados do presente trabalho evidenciam que o teste de avidez poderá ser mais um método valioso a ser utilizado no diagnóstico da toxoplasmose congênita em RNs.

Increasing number of secondary dengue cases: a concern

ackground: Dengue fever is caused by mosquito borne arbovirus of family Flaviviridae, Aedes agypti as the principle vector. In the recent past Delhi has witnessed several outbreaks affecting thousands of individuals and many of them get re-infected during subsequent years forming a bulk of secondary dengue cases putting them at risk of developing severe dengue.Methods: A total of 150 serum samples from suspected dengue cases were tested for dengue fever by NS-1 antigen and IgM antibody enzyme-linked immunosorbent assay (ELISA) followed by categorization into primary and secondary dengue using IgG avidity ELISA.Results: Out of total 150 clinically suspected dengue cases, 56 were positive either by Dengue NS-1 antigen or dengue IgM antibody or both. On the basis of dengue IgG avidity ELISA among 56 diagnosed dengue cases, 30 (53.57%) were found to be of secondary dengue.Conclusions: There is increasing trend of dengue cases in Delhi since past one decade. Being hyper-endemic area for dengue, more than 25% population have been reported to have past infection of dengue. Due to increased prevalence and simultaneous circulation of more than one serotypes, number of secondary dengue cases is also increasing. Since majority of severe dengue cases are associated with secondary dengue, early diagnosis and treatment can significantly reduce the fatal outcome. Thus, avidity testing for IgG antibody becomes an important tool.

Synthesis and evaluation of necrosis avidity of MRI contrast agent Gd-DO3A-Ether-Rhein / 中国药科大学学报

@#The aim of this study was to synthesize and evaluate the necrosis avidity of MRI contrast agent based on rhein and linked by ether. The novel ligand 10-{［6-(1, 8-dihydroxyanthraquinone-3-carboxamido)ethoxyethyl］amino}carbonylmethyl-1, 4, 7, 10-tetraazacyclododecan-1, 4, 7-triacetic acid(DO3A-Ether-Rhein, E1)was synthesized by two steps of acylation and deprotection reaction. The paramagnetic gadolinium 10-{［6-(1, 8-dihydroxyanthraquinone-3-carboxamido)ethoxyethyl］amino}carbonylmethyl-1, 4, 7, 10-tetraazacyclododecan-1, 4, 7-triacetic acid(Gd-DO3A-Ether-Rhein, GdE1)was obtained by coordination of Gd3+ with the above ligand. We examined the necrotic avidity of GdE1 in human hepatocellular carcinoma HepG2 cell necrosis induced by hyperthermia in vitro and in rat model with muscular necrosis induced by microwave ablation in vivo by MRI. The MRI was implemented before administration of GdE1 and during 0-9 h after administration of GdE1(0. 1 mmol/kg), and Gd-DOTA(gadolinium 1, 4, 7, 10-tetraacetic acid-1, 4, 7, 10-tetraazacyclo dodecane)was used as control. The signal intensity of necrotic cells(4 369±70)was significantly higher than that of normal cells(2 555±84)(P< 0. 05). Similarly, the contrast ratio between necrotic and normal muscle at 3 h after administration of GdE1(2. 00±0. 12)was remarkblely higher than that at 0 h after administration of GdE1(1. 27±0. 03)(P< 0. 05). Therefore, GdE1 presents good necrosis affinity and has the potential to be used in the diagnosis of necrosis-related diseases.

Effect of chaotropes in Chagas disease and leishmaniasis cross-reacting serology assays for epidemiological surveys

Abstract INTRODUCTION: Serological cross-reactivity between leishmaniasis and Chagas disease, especially at low titers, leads to difficulties of the seroepidemiological interpretation. METHODS: We have studied the ability of urea as a chaotrope to select high-avidity antibodies in IgG ELISA, thus reducing low-avidity IgG cross-reactivity in serologically positive samples in both assays. RESULTS: Using 0.5M urea for diluting the sample efficiently defined leishmaniasis or double infections in high-avidity IgG ELISA and eliminated false-positive results. CONCLUSIONS: The use of a chaotropic diluting agent is useful for improving the specificity of Chagas disease and leishmaniasis immunoassays.

Diagnostic significance of immunoglobulin G avidity in symptomatic and asymptomatic West Nile virus infection

Abstract INTRODUCTION West Nile virus (WNV) immunoglobulin M (IgM) antibodies have been shown to persist for up to 500 days in certain patients. To evaluate the usefulness of immunoglobulin G (IgG) avidity assessment in the diagnosis of WNV infection, we analyzed 54 WNV IgM- and/or IgG-positive serum samples from 39 patients with neuroinvasive disease and 15 asymptomatic cases tested during a seroprevalence investigation. METHODS Serological tests (WNV IgM/IgG antibody detection, IgG avidity) were performed using commercially available enzyme-linked immunosorbent assays. RESULTS WNV IgM antibodies were detected in 47 (87%) samples. Acute/recent WNV infection was confirmed based on low/borderline avidity index (AI) in 44 IgM-positive samples (93.6%). In three IgM-positive samples (6.4%), high IgG AIs were detected, thus indicating persisting IgM antibodies from previous infections. All IgM-negative samples showed high AIs. Patients with WNV neuroinvasive disease tested within 30 days showed low AIs. In six patients tested 34-50 days after disease onset, AI was borderline (42%-60%), suggesting earlier WNV IgG maturation. Samples with the highest IgM values were associated with the lowest AIs (Spearman's rho coefficient -0.767, p < 0.001). CONCLUSIONS Our results indicate that IgG avidity differentiates current/recent WNV infection from persistent IgM seropositivity from the previous WNV transmission season both in patients with WNV neuroinvasive disease and in asymptomatic persons. A strong negative correlation between IgM antibody levels and AI indicates that in cases with very high IgM levels, determination of IgG avidity may not be necessary. As many patients showed rapid avidity maturation, low IgG avidity is indicative of WNV infection within the previous month.

Estimating HIV incidence among female sex workers and injection drug users in Dehong Prefecture, 2009-2017 / 中华预防医学杂志

Objective@#To obtain HIV incidence among injection drug users (IDU) and female sex workers (FSW) in Dehong Prefecture, Yunnan Province during 2009-2017.@*Methods@#We recruited drug users and female sex workers from all sentinel surveillance sites across Dehong Prefecture during 2009-2017. A total of 10 480 IDU and 18 126 FSW in Dehong Prefecture were recruited by fingerprint technique. Data about drug uses, commercial sexual behavior, sociodemographic characteristics was collected by structured questionnaire. HIV-positive patients who were long-term infected or with CD4+ T cell count was ≤200 were not included for further HIV incidence testing. Also, those who self-identified as on antiretroviral treatment (ART) or AIDS cases were also excluded. A total of 841 and 157 plasma specimens from IDU and FSW that met the inclusion criterion were finally included, respectively. Limiting antigen avidity enzyme immunoassay(LAg-Avidity EIA) were performed to calculate the HIV incidence among these two sub-populations.@*Results@#A total of 3 444 IDU were HIV-positive, among which 884 (25.7%) were Burmese with age of (30.4±7.7), and 2 560 were Chinese with age of (36.6±7.3). Among 228 HIV-positive FSW, 109 (47.8%) were Burmese with age of (27.1±6.3), 119 (52.5%) were Chinese with age of (29.9±11.1). For IDU, the estimated HIV incidence among Burmese in 2009-2010, 2011-2012, 2013-2014, 2015-2017 was 4.20% (95%CI: -0.55%-8.95%), 7.75% (95%CI: 2.95%-12.55%), 11.79% (95%CI: 5.38%-18.20%), 10.30% (95%CI: 5.67%-14.94%), respectively, while Chinese were 3.11% (95%CI: 1.59%-4.64%), 0.03% (95%CI: -0.03%-0.08%), 1.55% (95%CI: 0.54%-2.57%), 0.58% (95%CI: -0.06%-1.04%), respectively. In 2009-2011, 2012-2014, 2015-2017, estimated HIV incidence among Burmese FSW was 0.22% (95%CI: -0.21%-0.64%), 1.24%(95%CI: 0.15%-2.32%), 0.55%(95%CI: 0.01%-1.08%). Whereas, estimated HIV incidence among Chinese FSW was 0.62% (95%CI: 0.25%-0.98%), 0.11% (95%CI: -0.04%-0.26%), 0.22% (95%CI: 0-0.44%).@*Conclusion@#HIV incidences among Chinese IDU and FSW are on the downward trend, while Burmese IDU and FSW seem to be gaining momentum.

The Predictive Values of Lesion Size, F-18 FDG Avidity and I-131 Avidity for the Clinical Outcome of I-131 Treatment in Patients with Metastatic Differentiated Thyroid Carcinoma Only in the Lung / 대한핵의학회잡지

PURPOSE: We aimed to evaluate the prognostic values of radiography, F-18 FDG PET, and I-131 whole body scans in patients with lung-only metastasis from differentiated thyroid carcinoma (DTC).METHODS: Between 1998 and 2013, we included 31 patients (F: 26, M: 5) with lung-only metastasis from DTC who had been treated with I-131 and underwent PET. Lung metastasis was categorized according to the size (macronodular ≥1.0 cm vs. micronodular <1.0 cm), FDG avidity (avid vs. non-avid), and I-131 avidity (avid vs. non-avid). Progression-free survival (PFS) was evaluated for each patient.RESULTS: Among 31 patients, seven (23%) had macronodular lung metastasis, 26 (84%) had FDG avid lung metastasis, and 16 (52%) had I-131 avid lung metastasis. During the median follow-up period of 9.4 y, median PFS was 6.1 y. Based on Kaplan-Meier analysis, macronodular lung metastasis (p = 0.017) and I-131 non-avid lung metastasis (p = 0.059) were significantly associated with worse outcomes, but FDG avid lung metastasis was not (p = 0.135). Patients with FDG non-avid lung metastasis did not experience disease progression during follow-up, while 11 of 26 patients (42%) experienced disease progression. Based on univariate analysis, the hazard ratio for a poor prognosis was 3.78 (p = 0.029) for macronodular lung metastasis and 3.29 (p = 0.079) for I-131 non-avid lung metastasis.CONCLUSIONS: Macronodular and I-131 non-avid lung metastasis were associated with a poor prognosis in lung-only metastasis from DTC. Although FDG avid lung metastasis may be associated with a poor prognosis, a larger-scale study is needed.

Serologic Tests of IgG and IgM Antibodies and IgG Avidity for Diagnosis of Ocular Toxoplasmosis

This prospective study was aimed to detect acute and chronic ocular toxoplasmosis by comparison of anti-Toxoplasma gondii IgM and IgG antibody levels and IgG avidity test. One hundred and seventeen patients with ocular toxoplasmosis (OT) who referred to the Farabi Eye Hospital, Tehran, Iran were included in this study. Of the patients, 77 cases were positive for anti-T. gondii IgG, and 8 cases were positive for anti-T. gondii IgM. IgG avidity test revealed 11, 4, and 102 cases were low, intermediate, and high, respectively, and 6.8% and 9.4% of cases were positive for IgM and IgG avidity tests, respectively (P=0.632). Agreement (Kappa value) between paired tests IgG-IgM, IgG-IgG avidity, and IgM-IgG avidity was 0.080, 0.099, and 0.721, respectively (P < 0.05). This study showed that conventional serologic tests (IgM and IgG levels) and IgG avidity correlate well each other and can be used to differentiate recent infections from old OT. It seems that reactivated old infections rather than recently acquired infections are majority of Iranian OT patients.

A Case of Anti-JMH Antibody with High-Titer, Low-Avidity Characteristics: The First Case in Korea / 대한수혈학회지

Anti-John Milton Hagen (JMH) is a high-titer, low-avidity (HTLA) antibody against the high frequency red blood cell (RBC) antigen JMH. It occurs very rarely and has not yet been reported in Korea. Here, we report a case of anti-JMH antibody identified in a 92-year-old man without previous blood transfusion history, who had been hospitalized with pneumonia. The patient's hemoglobin level was reduced to 7.6 g/dL on the 35th day of hospitalization, requiring RBC transfusion. Antibody identification test revealed antibodies that showed pan-reactivity to all panel cells at the antiglobulin phase. A titration test confirmed that it was a HTLA antibody. He was given one least-incompatible unit of RBC without any adverse events, and his hemoglobin level increased to 9.3 g/dL. The patient's sample was referred to a reference laboratory and the antibody was identified as anti-JMH. He was successfully transfused with 6 additional units of least-incompatible RBCs without complication. HTLA antibodies against high frequency antigens, such as anti-JMH, are less likely to cause significant destruction of transfused antigen positive RBCs. However, identifying the specificity of these antibodies is necessary to appropriately understanding the clinical significance of the antibody, detecting other clinically important alloantibodies that may coexist, and determining the appropriate blood for transfusion.

Passive acquisition of anti-Staphylococcus aureus antibodies by newborns via transplacental transfer and breastfeeding, regardless of maternal colonization

OBJECTIVE: To investigate the transmission of anti-Staphylococcus aureus (Sa) IgG, IgG1 and IgG2 via placental transfer and the transfer of IgA via the colostrum according to maternal Sa carrier status at delivery. METHODS: We evaluated anti-Sa IgG, IgG1 and IgG2 in maternal and cord sera and IgA in colostrum from a case (n=49, Sa+) and a control group (n=98, Sa-). RESULTS: Of the 250 parturients analyzed for this study, 49 were nasally colonized with S. aureus (prevalence of 19.6%). Ninety-eight non-colonized subjects were selected for the control group. The anti-Sa IgG, IgG1 and IgG2 levels and the IgG avidity indexes in the maternal and cord sera did not differ between the groups, with a low transfer ratio of anti-Sa IgG to the newborns in both groups. The anti-Sa IgG2 titers were significantly higher than the IgG1 titers in the maternal and cord sera. Inversely, the transfer ratios were higher for anti-Sa IgG1 compared with IgG2; however, no differences between the groups were detected. The Sa-specific IgA levels and avidity indexes in the colostrum were equivalent between groups. CONCLUSIONS: Maternal Sa nasal colonization at delivery is not associated with higher antibody levels in the mother or newborns. The high titers of anti-Sa IgG2 found in the cord serum indicate a greater reactivity with non-protein antigens, which may further contribute to the susceptibility to staphylococcal infections at birth. The presence of IgA in the colostrum with avidity to S. aureus reinforces the importance of breastfeeding shortly after birth.

Kinetics and avidity of anti-Toxocara antibodies (IgG) in rabbits experimentally infected with Toxocara canis / Cinética e avidez de anticorpos (IgG) anti-Toxocara em coelhos experimentalmente infectados com Toxocara canis

Abstract An evaluation was made of the kinetics and avidity of anti-Toxocara antibodies (IgG) in rabbits experimentally infected with embryonated Toxocara canis eggs. Seventeen four month old New Zealand White rabbits were distributed into two groups. In the experimental group, twelve rabbits were infected orally with 1,000 embryonated T. canis eggs. A second group (n = 5), uninfected, was used as a control. Serum samples were collected for analysis on days 7, 14, 21, 28 and 60 post-infection (DPI). An indirect ELISA test was performed to evaluate the reactivity index (RI) of IgG anti-T. canis antibodies and to calculate the avidity index (AI). The animals showed seroconversion from the 14th DPI, with high AI (over 50%) except for one animal, which presented an intermediate AI. At 60 DPI, all the animals were seropositive and maintained a high AI. The data indicated that specific IgG antibodies formed early (14 DPI) in rabbits infected with T. canis, with a high avidity index that persisted throughout the course of the infection.

Resumo O objetivo deste estudo foi o de avaliar a cinética e a avidez de anticorpos anti-Toxocara canis, em coelhas infectadas experimentalmente com ovos embrionados de Toxocara canis. Foram utilizados 17 coelhos New Zealand de linhagem branca, com quatro meses de idade, distribuídos em dois grupos. No grupo experimental, doze coelhas foram infectadas, oralmente, com 1.000 ovos larvados de T. canis. Um segundo grupo (n=5), não infectado, foi utilizado como controle. Nos dias 7, 14, 21, 28 e 60 pós-infecção (DPI), foram coletadas amostras de soro para análise. O teste de ELISA indireto foi realizado para avaliar o índice de reatividade (IR) de anticorpos IgG anti-T. canis e para cálculo do índice de avidez (IA). A soroconversão nos animais ocorreu a partir do140 DPI, com verificação de alto IA (superior a 50%), com exceção de um animal, que apresentou médio IA. Aos 60 DPI, todos os animais foram soropositivos e mantiveram alto IA. Os dados mostram que em coelhos infectados por T. canis, anticorpos IgG específicos formam-se precocemente (14 DPI), apresentando alto índice de avidez e que se mantém durante o curso da infecção.

Na boca do crocodilo: a face indigesta da amamentação exclusiva / In the crocodile's mouth: the indigestible face of exclusive breastfeeding

O presente artigo problematiza as possíveis implicações dos programas de aleitamento materno exclusivo, que, fortemente firmados em um ideal de maternidade, garantem o cumprimento de prescrições reguladas pela chamada livre-demanda, devendo a mãe oferecer ao bebê, pelo menos até o sexto mês, somente o seio-leite materno como alimento e objeto oral. Retomando algumas considerações de Jacques Lacan, propomos pensar na tendência a uma excessiva presença da mãe ensejada por esse discurso que visa à benevolência, fazendo surgir, como possível contraponto, o bebê enquanto um objeto-resto na fantasia materna. Poderíamos pensar o discurso do aleitamento como favorecendo um prolongamento da relação alienante entre mãe e filho, com graves consequências para o bebê? No âmbito da fantasia materna, poderia o bebê ser tomado como um objeto de gozo para o Outro? São questões a serem desdobradas ao longo do texto.

This article discusses the possible implications of the exclusive breastfeeding programs which, strongly grounded on an ideal of motherhood, ensure the fulfillment of prescriptions regulated by the breastfeeding on demand, ruling that the mother should just provide to the baby, at least until the sixth month, the breast-milk as the only food and oral object. Resuming some considerations of Jacques Lacan, we propose to think of the tendency to excessive presence of the mother occasioned by this discourse that aims to benevolence, doing emerge, as a possible counterpoint, the baby like a rest-object at the maternal fantasy. Could we think the discourse of breastfeeding as favoring a prolongation of alienating relationship between mother and child, with serious consequences for the baby? As regards the maternal fantasy, the baby could be taken as an object of the jouissance for the Other? These are issues to be discussed throughout the text.

Comparison of immunofluorescence and enzyme-linked immunosorbent assay and immunoglobulin G avidity techniques for screening of anti: Toxoplasma antibodies among single serum sample pregnant women in Tabriz, Iran.

Background: Congenital toxoplasmosis is that pregnant women acquire the infection during gestation; diagnosis of the acute infection during pregnancy is a complex subject of maternal toxoplasmosis. Thus, the presence of immunoglobulin G (IgG) and/or IgM Toxoplasma antibodies in a single serum sample drawn during gestation cannot be used to defi ne whether the infection was recently acquired or chronic. Materials and Methods: At this cross-sectional descriptive study, sera of 391 pregnant women examined and compared. They were in an age range of 21-35 years, referred by gynecologists and infectious disease specialists, during March 2012-April 2013. They have referred, 215 (54.98%), 102 (26%), 74 (18.92%) in the fi rst, second and third trimesters of gestation, respectively. For each of them, a questionnaire was completed and serum samples were prepared in an equal condition, examined according to the procedures of indirect immunofl uorescence (IIF), enzyme-linked immunosorbent assay (ELISA) and IgG Avidity techniques. Results: We have found 111 (28.38%) seronegative and 280 (71.61%) seropositive cases by IIF and 124 (31.70%) seronegative, 267 (68.28%) seropositive cases by ELISA. The IgG avidity test confi rmed 45 (69.23%) and 7 (10.76%) doubtful cases of IgM test in IIF and ELISA techniques. Conclusions: This study highlights how to manage pregnant women with toxoplasmosis, especially in a single serum sample condition.

Occurrence of anti-Toxoplasma gondii antibodies in meat and dairy goat herds in Rio Grande do Norte, Brazil / Ocorrência de anticorpos anti-Toxoplasma gondii em rebanhos caprinos de corte e leiteiros do Rio Grande do Norte, Brasil

Toxoplasmosis is caused by Toxoplasma gondii, which is the main causative agent of abortion in small ruminants. Goats are among the animals that are most susceptible to this protozoon, and the disease that it causes leads to significant economic losses and has implications for public health, since presence of the parasite in products of goat origin is one of the main sources of human infection. Because of the significant economic impact, there is an urgent need to study the prevalence of T. gondii infection among goats in Sertão do Cabugi, which is the largest goat-producing region in Rio Grande do Norte. In the present study, the ELISA assay was used to test 244 serum samples from nine farms, located in four different municipalities in the Sertão do Cabugi region, which is an important goat-rearing region. The results showed that the prevalence of anti-T. gondii antibodies was 47.1% and that there was a significant association between positivity and the variables of age (≥ 34 months), location (Lajes, Angicos and Afonso Bezerra) and farm (all the farms). The avidity test was applied to all the 115 ELISA-positive samples to distinguish between acute and chronic infection. One hundred and three samples (89.6%) displayed high-avidity antibodies, thus indicating that most of the animals presented chronic infection, with a consequent great impact on the development of the goat production system and a risk to human health.

A toxoplasmose é causada pelo Toxoplasma gondii, principal agente causador de aborto em pequenos ruminantes. Os caprinos são uns dos animais mais suscetíveis a esse protozoário, levando a perdas econômicas significativas e implicações para a saúde pública, uma vez que a presença do parasito em produtos de origem caprina é uma das principais fontes de infecção humana. Devido ao impacto econômico significativo torna-se urgente estudar a prevalência da infecção, pelo T. gondii, entre caprinos do Sertão do Cabugi, a maior região produtora de caprinos no Rio Grande do Norte. O presente estudo utilizou o ELISA para testar 244 amostras de soro de 9 fazendas, situadas em 4 diferentes cidades na região do Sertão do Cabugi; uma importante região de criação de cabras. Os resultados mostraram uma prevalência de 47,1% para anticorpos anti- T. gondii e uma significativa associação entre a positividade e as variáveis idade (≥ 34 meses), localização (Lajes, Angicos e Afonso Bezerra e propriedade (todas as fazendas). O teste de avidez foi aplicado a todas as 115 amostras positivas pelo ELISA para discriminar entre infecção aguda e crônica. Cento e três amostras (89,6%) apresentaram anticorpos de alta avidez; indicando que a maioria dos animais estavam em infecção crônica, gerando um grande impacto sobre o desenvolvimento do sistema de produção em cabras e um risco para a saúde humana.

Seroprevalencia y detección de infección primaria por citomegalovirus mediante prueba de avidez IgG en el primer trimestre de embarazo / Seroprevalence and detection of primary infection by cytomegalovirus with IgG avidity test during the first quarter of pregnancy

Objetivo. Conocer la seroprevalencia y detección de infección primaria por citomegalovirus (CMV) mediante prueba de avidez de inmunoglobulina G (IgG) durante el primer trimestre del embarazo en el Hospital General de Morelia, Michoacán. Material y métodos. Se estudiaron 177 pacientes mediante prueba de Elisa modificada, la cual utiliza inmunoanálisis quimioluminiscente de micropartículas (CMIA) para detección de anti-CMV (IgG e inmunoglobulina M [IgM]) e IgG avidez. Resultados. Del total de pruebas, 90.4% resultaron positivas para IgG; de éstas, 2.3% resultaron reactivas a IgM. En este segundo grupo, la prueba de IgG avidez reportó avidez baja en 1.1% y alta en el mismo porcentaje; 9.6% fueron seronegativas. Conclusiones. Se encontró similitud con lo publicado en México. Los profesionales de la salud deben conocer los algoritmos para el diagnóstico y manejo oportuno de la infección por CMV mediante la prueba de avidez de IgG.

Objective. To determine the seroprevalence and detection of primary infection by cytomegalovirus (CMV) with immunoglobulin G (IgG) avidity test during the first quarter of pregnancy in the General Hospital in Morelia, Michoacan. Materials and methods. A total of 177 patients were studied employing a modified Elisa test using a chemiluminescent microparticle immunoassay (CMIA) for the detection of CMV antibodies (IgG and immunoglobulin M [IgM]), and IgG avidity. Results. 90.4% were positive for IgG, and of these, 2.3% were also reactive for IgM, and in this group the IgG avidity test reported low avidity for 1.1% and higher avidity in the same percentage. 9.6% were seronegative. Conclusions. Similarity was found with published studies in Mexico. Health professionals should know the clinical algorithms for diagnosis and proper management of CMV infection using the IgG avidity test.

High Expression of Water-Soluble Recombinant Antigenic Domains of Toxoplasma gondii Secretory Organelles

Recombinant antigenic proteins of Toxoplasma gondii are alternative source of antigens which are easily obtainable for serodiagnosis of toxoplasmosis. In this study, highly antigenic secretory organellar proteins, dense granular GRA2 and GRA3, rhoptrial ROP2, and micronemal MIC2, were analyzed by bioinformatics approach to express as water-soluble forms of antigenic domains. The transmembrane region and disorder tendency of 4 secretory proteins were predicted to clone the genes into pGEX-4T-1 vector. Recombinant plasmids were transformed into BL21 (DE3) pLysS E. coli, and GST fusion proteins were expressed with IPTG. As a result, GST fusion proteins with GRA225-105, GRA339-138, ROP2324-561, and MIC21-284 domains had respectively higher value of IgG avidity. The rGST-GRA225-105 and rGST-GRA339-138 were soluble, while rGST-ROP2324-561 and rGST-MIC21-284 were not. GRA231-71, intrinsically unstructured domain (IUD) of GRA2, was used as a linker to enhance the solubility. The rGST-GRA231-71-ROP2324-561, a chimeric protein, appeared to be soluble. Moreover, rGST-GRA231-71-MIC21-284 was also soluble and had higher IgG avidity comparing to rGST-MIC21-284. These 4 highly expressed and water-soluble recombinant antigenic proteins may be promising candidates to improve the serodiagnosis of toxoplasmosis in addition to the major surface antigen of SAG1.

IgG Avidity Antibodies against Toxoplasma gondii in High Risk Females of Reproductive Age Group in India

Toxoplasma gondii is an obligate intracellular protozoan that is distributed worldwide. Recently, several tests for avidity of Toxoplasma IgG antibodies have been introduced to help discriminate between recently acquired and distant infections. The study was conducted in Jawaharlal Nehru Medical College and Hospital, India from February 2011 to September 2012. Serum specimens were subjected to Toxoplasma IgM ELISA and IgG avidity ELISA test. Out of 48 patients with abortions, 17 (35.4%) were positive for IgM ELISA, and 8 (16.6%) had low IgG avidity antibodies. Out of 48 patients with other obstetric problems, 23 (47.9%) were positive for IgM ELISA, and 17 (35.4%) had low IgG avidity antibodies. Combining both groups on avidity test, only 25 of 40 (62.5%) IgM-positive women had low-avidity IgG antibodies suggesting a recent T. gondii infection in these women. More importantly, 15 (37.5%) of the IgM-positive women had high-avidity antibodies suggesting that the infection was acquired before gestation The relation of IgM seropositivity with the following risk factors was not found to be statistically significant; contact with cats (0.13), non-vegetarian food habits (0.05), and low socio-economic status (0.49). While, for IgG avidity ELISA, only contact with cats (0.01) was significantly associated with seropositivity. All other risk factors have P-values of >0.05 (not significant). IgG avidity test when used in combination with IgM test was a valuable assay for diagnosis of ongoing or recently acquired T. gondii infection in India.

IgG AVIDITY WESTERN BLOT USING Toxoplasma gondii rGRA-7 CLONED FROM NUCLEOTIDES 39-711 FOR SERODIAGNOSIS OF ACUTE TOXOPLASMOSIS / Avidez do IgG pelo Western Blot usando o r GRA-7 do Toxoplasma gondii clonado de nucleotídeos 39-711 para sorodiagnóstico de toxoplasmose aguda

Toxoplasmosis is an important cause of congenital infection. The present study was performed to evaluate the usefulness of recombinant (r) GRA-7 cloned from nucleotides (n) 39-711 in discriminating between acute and chronic toxoplasmosis. First, commercial IgM, IgG and IgG avidity ELISAs were used to determine the serological profile of the sera. Serum samples were from 20 symptomatic patients with acute infection (low IgG avidity, IgM positive), 10 with chronic infection (high IgG avidity, IgM negative) and 10 with indeterminate IgG avidity (IgM positive) which were tested for IgG avidity status with an in-house developed IgG avidity Western blot using the rGRA-7 recombinant antigen. All 20 sera from cases of probable acute infection showed bands which either faded out completely or reduced significantly in intensity after treatment with 8 M urea, whereas the band intensities of the 10 serum samples from chronic cases remained the same. Of the 10 sera with indeterminate IgG avidity status, after treatment with 8 M urea the band intensities with six sera remained the same, two sera had completely faded bands and another two sera had significantly reduced band intensities. Discrimination between acute and chronic toxoplasmosis was successfully performed by the in-house IgG avidity Western blot.

Toxoplasmose é uma causa importante de infecção congênita. O presente estudo foi feito para avaliar o uso do recombinante (r) GRA-7 clonado de nucleotídeos (n) 30-711 para discriminar entre toxoplasmose aguda e crônica. Inicialmente IgM, IgG e ELISA avidez IgG comerciais foram usados para determinar o perfil sorológico do soro. Amostras de soro de 20 pacientes sintomáticos com infecção aguda (IgG avidez baixa, IgM positivo), 10 com infecção crônica (alta avidez IgG, IgM negativo) e 10 com avidez IgG indeterminada (IgM positivo) que foram testados para o status de avidez IgG com um doméstico Western Blot desenvolvendo avidez IgG usando o rGRA-7 antígeno recombinante. Todos os 20 soros de provável infecção aguda mostraram bandas que ou se apagaram completamente ou tiveram a sua intensidade significantemente reduzida após tratamento com uréia 8 M, enquanto as intensidades das bandas das 10 amostras de soros de casos crônicos permaneceram iguais. Dos 10 soros com status indeterminado de avidez de IgG, após tratamento com uréia 8 M a intensidade das bandas em seis soros permaneceram iguais, dois soros tiveram bandas apagadas completamente e dois outros tiveram significante redução da intensidade das bandas. Discriminação entre toxoplasmose aguda e crônica foi feita com sucesso através do IgG avidez Western blot doméstico.

Case-control study of an outbreak of acute toxoplasmosis in an industrial plant in the state of São Paulo, Brazil / Estudo caso-controle de surto de toxoplasmose aguda em indústria no estado de São Paulo, Brasil

Foodborne diseases represent operational risks in industrial restaurants. We described an outbreak of nine clustered cases of acute illness resembling acute toxoplasmosis in an industrial plant with 2300 employees. These patients and another 36 similar asymptomatic employees were diagnosed with anti-T. gondii IgG titer and avidity by ELISA. We excluded 14 patients based on high IgG avidity and chronic toxoplasmosis: 13 from controls and one from acute disease other than T. gondii infection. We also identified another three asymptomatic employees with T.gondii acute infection and also anti-T. gondii IgM positive as remaining acute cases. Case control study was conducted by interview in 11 acute infections and 20 negative controls. The ingestion of green vegetables, but not meat or water, was observed to be associated with the incidence of acute disease. These data reinforce the importance of sanitation control in industrial restaurants and also demonstrate the need for improvement in quality control regarding vegetables at risk for T. gondii oocyst contamination. We emphasized the accurate diagnosis of indexed cases and the detection of asymptomatic infections to determine the extent of the toxoplasmosis outbreak.

Doenças transmitidas por alimentos representam riscos operacionais em restaurantes industriais. Descrevemos surto de nove casos agrupados de doença aguda semelhante à toxoplasmose em indústria de 2300 funcionários. Estes pacientes e outros 36 funcionários assintomáticos foram diagnosticados por ELISA para o título e avidez de IgG anti-T. gondii. Foram excluídos 14 pacientes com toxoplasmose crônica e alta avidez: 13 de controles e um de doença aguda não relacionada à infecção por T. gondii. Também identificamos três empregados assintomáticos com infecção aguda por T.gondii, que como os restantes agudos apresentavam anti-T.gondii IgM ELISA positivo. Conduzimos estudo caso controle por entrevista em 11 infecções agudas e 20 controles negativos. A ingestão de vegetais, mas não de carne ou água, foi associada com a incidência da doença aguda. Esses dados reforçam a importância do controle sanitário em restaurantes industriais e também demonstram a necessidade de melhoria no controle de qualidade sobre vegetais em risco de contaminação por oocistos de T. gondii. Enfatizamos o diagnóstico preciso de casos e a detecção de infecções assintomáticas para determinar a extensão do surto de toxoplasmose.