Serological evaluation of the schistosome's secretory enzyme phytochelatin synthase and phosphoglycerate mutase for the detection of human Schistosoma japonicum infection.

Secretory enzymes from Schistosoma japonicum are promising candidate antigens in the diagnosis of schistosomiasis. Our previous studies have proven that thioredoxin peroxidase-1 (SjTPx-1) is useful for the detection of this parasitic disease in humans, water buffaloes, and dogs. In this study, we evaluated two more secretory enzymes namely phosphoglycerate mutase (SjPGM) and phytochelatin synthase (SjPCS) with SjTPx-1 as the reference antigen. SjPGM was shown to have good diagnostic potentials in animal samples in previous studies, whereas SjPCS was chosen because of its absence in the mammalian hosts. Serum samples including 96 endemic negative controls, 107 schistosomiasis japonica positive samples, and 31 samples positive for other parasitic trematode infections (Clonorchis sinensis, Opisthorchis viverrini, Paragonimus westermani) were tested with the antigens using enzyme-linked immunosorbent assay. Results showed that SjPCS detected more positive samples and had fewer cross-reactions than SjPGM. With 85.05% sensitivity and 93.55% specificity, SjPCS can therefore be used in the detection of human schistosomiasis.

Evaluation of crude adult Ascaris suum intestinal tract homogenate in inducing protective IgG production against A. suum larvae in BALB/c mice.

Globally, ascariasis ranks as the second leading intestinal helminth infection. However, progress in developing better control strategies, such as vaccines, remains slow-paced. This study aims to measure antibody production and parasite load in male BALB/c mice immunized with crude Ascaris suum intestinal tract homogenate. Thirty-two (32) mice were randomized into: (1) unvaccinated, uninfected (UU); (2) unvaccinated, infected (UI); (3) vaccinated, uninfected (VU); and (4) vaccinated, infected (VI) groups. A 100-µL vaccine containing 50 µg of homogenized A. suum intestines and Complete Freund's Adjuvant (1:1) were introduced intraperitoneally. Immunizations were done on days 0, 10, and 20. Oral gavage with 1000 embryonated eggs was done on day 30. Blood was obtained at day 40. To measure serum IgG levels, indirect ELISA was done. Microtiter plates were coated with 100 µg larval homogenate, and HRP-conjugated anti-mouse IgG was used as secondary antibody. Parasite load was measured in lung and liver tissues. Tukey's HSD of signal to cut-off ratios of absorbance readings obtained in indirect ELISA procedure for the 1:200 serum dilution showed statistically significant difference between the UU and VI (p = 0.026) as well as between UI and VI (p = 0.003) groups. No statistically significant difference in parasite load was observed in the lungs (p = 0.074), liver (p = 0.130), and both lungs and liver (p = 0.101). Immunization elicited a significant larva-directed IgG production. However, there is no significant difference in parasite loads in either lung or liver tissues across all treatment groups as the larval counts obtained from the study were very low and may not be indicative of the actual parasite load in mice.

Integrative taxonomic approach of trypanosomes in the blood of rodents and soricids in Asian countries, with the description of three new species.

Trypanosoma lewisi (Kinetoplastea: Trypanosomatida: Trypanosomatidae) with a cosmopolitan distribution is the type species of the subgenus Herpetosoma, which includes ca. 50 nominal species isolated mainly from rodents. Since members of Herpetosoma in different host species have an almost identical morphology of bloodstream forms, these trypanosomes are referred to as 'T. lewisi-like', and the molecular genetic characterization of each species is necessary to verify their taxonomy. In the present study, we collected blood samples from 89 murid rodents of 15 species and 11 soricids of four species in Indonesia, Philippines, Vietnam, Taiwan, and mainland China for the detection of hemoprotozoan infection. T. lewisi and T. lewisi-like trypanosomes were found in the blood smears of 10 murid animals, which included Bandicota indica (two rats), Rattus argentiventer (one rat), and Rattus tiomanicus (two rats) in Indonesia; Rattus rattus (one rat) in the Philippines; and Niviventer confucianus (four rats) in mainland China. Furthermore, large- or medium-sized non-T. lewisi-like trypanosomes were detected in two soricids, Crocidura dracula in Vietnam and Anourosorex yamashinai in Taiwan, respectively. Molecular genetic characterization of the small subunit (SSU) ribosomal RNA gene (rDNA) and glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) gene indicated that the trypanosomes from all the murid hosts had identical SSU rDNA or gGAPDH gene nucleotide sequences except for those in N. confucianus in mainland China. These N. confucianus-infecting trypanosomes also showed several unique morphological features such as smaller bodies, anteriorly positioned nuclei, and larger rod-shaped kinetoplasts when compared with T. lewisi trypomastigotes. Trypanosoma (Herpetosoma) niviventerae n. sp. is erected for this new species. Similarly, based on morphological and molecular genetic characterization, Trypanosoma sapaensis n. sp. and Trypanosoma anourosoricis n. sp. are proposed for the trypanosomes in C. dracula in Vietnam and A. yamashinai in Taiwan, respectively. More effort directed toward the morphological and molecular genetic characterization of the trypanosomes of rodents and soricids is required to fully understand the real biodiversity of their hemoflagellates.

Schistosoma japonicum cathepsin B as potential diagnostic antigen for Asian zoonotic schistosomiasis.

In this study, the diagnostic value of Schistosoma japonicum cathepsin B (SjCatB) was evaluated as an antigen for the early detection of S. japonicum infection. SjCatB is a key protease used by the cercaria to penetrate the intact skin of the host for transdermal infection. The early exposure of the host's immune system to this enzyme may elicit early production of antibodies against this molecule. Therefore, the recombinant SjCatB (rSjCatB) was expressed in Escherichia coli with N-terminal 6xHis-tag. rSjCatB was tested for its performance as a diagnostic antigen using indirect enzyme-linked immunosorbent assay (ELISA) with sera from experimentally infected mice collected at > 8 weeks post-infection. Showing 100% sensitivity and 95.0% specificity in the ELISA, rSjCatB was then evaluated with sera from experimentally infected mice collected at 1-7 weeks post-infection to determine how early the antibodies can be detected. Results showed that as early as 6 weeks post-infection, 2 of the 3 infected mice were found to be positive with the antibodies against SjCatB. Furthermore, the potential of the recombinant antigen in detecting human schistosomiasis was evaluated with archived serum samples collected from individuals who had been diagnosed with S. japonicum infection by stool examination. Results showed 86.7% sensitivity and 96.7% specificity suggesting its high diagnostic potential for human schistosomiasis. In addition, SjCatB showed minimal cross-reaction with the sera collected from patients with other parasitic diseases. In conclusion, the results of this study suggest that SjCatB will be useful in the development of a sensitive and specific early detection test for S. japonicum infection.

Development and optimization of cocktail-ELISA for a unified surveillance of zoonotic schistosomiasis in multiple host species.

The zoonotic characteristic of the human parasite Schistosoma japonicum infecting a significant number of wild and domestic animals highlights the need to develop a unified surveillance in multiple host species for a strengthened schistosomiasis control. It has been shown in several studies that water buffaloes and dogs are considered important reservoirs in the transmission of the schistosome parasite to humans. Recombinant antigens like thioredoxin peroxidase-1 (SjTPx-1) and tandem repeat proteins (Sj1TR, Sj7TR) have been shown to be good diagnostic antigens individually in humans, water buffaloes, and dogs in previous studies. Mixing these antigens together in a cocktail-ELISA might not only improve their diagnostic potentials but rather produce a multi-host species detection means for zoonotic schistosomiasis. In this study, we aimed to develop and optimize cocktail-ELISA by testing different combinations of these recombinant antigens in humans, water buffaloes, and dogs. As compared with the diagnostic potential calculated for each of the three recombinant antigens used, their combination has presented improved specificities, positive predictive values, and kappa values. Using samples collected from various endemic areas in the Philippines, results showed that the combination of SjTPx-1/Sj7TR/Sj1TR has the highest sensitivity in humans (84.1 %), water buffaloes, and dogs (80 %) and specificity (100 %) in all host species. This study therefore suggests the use of cocktail-ELISA in improving the zoonotic surveillance in schistosomiasis endemic areas.

Determining the active role of microscopists in community awareness-raising activities for malaria prevention: a cross-sectional study in Palawan, the Philippines.

BACKGROUND: Malaria remains one of the most prevalent and fatal diseases among the inhabitants of Palawan in the Philippines. Palawan, where healthcare services remain limited, has the highest malaria endemicity in the country. To eliminate malaria, effective prevention measures should be conducted alongside early diagnosis and prompt treatment, which are the major tasks of the trained microscopists in Palawan. However, while the microscopists have implemented community awareness-raising activities aimed at preventing transmission of malaria, the nature and quality of these activities have not been evaluated. The present study identified the factors associated with the strengthening of community awareness-raising activities for malaria prevention implemented by microscopists in Palawan. METHODS: A cross-sectional study was conducted among 127 microscopists in Palawan. Data were collected using self-administered questionnaires from November 2010 to February 2011. For data analysis, structural equation modelling was conducted, based on the questionnaire results, to identify the impact of factors associated with the number of community malaria awareness-raising activities implemented by microscopists using the following assessment indicators: (1) place of assignment; (2) annual parasite index; (3) microscopists' capacity (service quality, knowledge on malaria, and ability in malaria microscopy); (4) self-preventive measures against malaria; and (5) job satisfaction. RESULTS: High microscopists' capacity was found to be a significant factor for a greater number of community awareness-raising activities for malaria prevention. High microscopists' capacity was significantly explained by its two sub-components: high service quality (active detection, diagnosis and treatment, prescription of anti-malarial, and follow-up) and high ability in malaria microscopy (preparation and documentation, slide preparation and observation, safe handling and disposal, and knowledge on the morphology of infected red blood cells). CONCLUSIONS: Microscopists' capacity was identified as a significant factor in community awareness-raising activities for malaria prevention. Thus, the strengthening of service quality and ability in malaria microscopy should be of the highest priority.

Malarial Antibody Detection with an Engineered Yeast Agglutination Assay.

Malaria, a disease caused by the Plasmodium parasite carried by Anopheles mosquitoes, is commonly diagnosed by microscopy of peripheral blood smears and with rapid diagnostic tests. Both methods show limited detection of low parasitemia that may maintain transmission and hinder malaria elimination. We have developed a novel agglutination assay in which modified Saccharomyces cerevisiae cells act as antigen-displaying bead-like particles to capture malaria antibodies. The Epidermal Growth Factor-1 like domain (EGF1) of the Plasmodium falciparum merozoite surface protein-1 (PfMSP-119) was displayed on the yeast surface and shown to be capable of binding antimalaria antibodies. Mixed with a second yeast strain displaying the Z domain of Protein A from Staphylococcus aureus and allowed to settle in a round-bottomed well, the yeast produce a visually distinctive agglutination test result: a tight "button" at a low level of malarial antibodies, and a diffuse "sheet" when higher antibody levels are present. Positive agglutination results were observed in malaria-positive human serum to a serum dilution of 1:100 to 1:125. Since the yeast cells are inexpensive to produce, the test may be amenable to local production in regions seeking malaria surveillance information to guide their elimination programs.

Entamoeba histolytica infections in a slum community in Manila, Philippines as detected by stool ELISA.

This study aimed to determine the prevalence of Entamoeba histolytica in BASECO, an urban slum community situated in Manila Harbor, Manila, Philippines using stool enzyme-linked immunosorbent assay (ELISA). It also aimed to determine if age, sex, and geographic location are contributory factors to the prevalence of E. histolytica. Stool samples were collected from 627 urban slum community residents of BASECO. Samples were viewed under light microscopy and the different parasites observed were identified. Stool ELISA was done using E. histolytica II antigen detection kits (TECHLAB®). Using E. histolytica II kits, E. histolytica had a prevalence of 9.09% (5/55) among the microscopically-positive samples for E. histolytica/E. dispar indicating a greater prevalence for the nonpathogenic species. No significant difference was observed in the prevalence of infection across all three variables: age, sex and geographic location. The overall prevalence of E. histolytica in BASECO, Manila, Philippines is 0.797% (5/627) which is lower than previous studies done on estimating the prevalence of E. histolytica using various techniques.

Factors determining job satisfaction of malaria community microscopists in Palawan, the Philippines: a cross-sectional mixed-methods study.

BACKGROUND: This study was conducted to investigate the overall job satisfaction and its associated factors among microscopists as community health workers who specialize in malaria microscopic diagnosis and treatment in Palawan, the Philippines. The study offered new insights to further lessen the burden of malaria and improve treatment by enhancing their commitment and satisfaction. METHODS: A quantitative study was conducted with 217 microscopists who were asked about their sociodemographic characteristics, ability as microscopists (service quality, knowledge of malaria and ability to diagnose malaria by microscopy [malaria microscopy]) and job satisfaction. Structural equation modelling (SEM) was conducted to analyse these factors. A qualitative study was also conducted and four focus group discussions (FGDs) were carried out. RESULTS: SEM identified that the job satisfaction of microscopists was enhanced by ability in malaria microscopy, the annual parasite index and belonging to ethnic minorities. In the FGDs, high job satisfaction was seen in their devotion to the community and increasing knowledge about malaria. CONCLUSIONS: Providing opportunities to improve malaria microscopy ability would increase the overall job satisfaction of microscopists and eventually improve the quality of care.

Satisfaction of Malaria Patients with Care by Community Health Workers Trained in Microscopy: A Mixed-Methods Study in Palawan, the Philippines.

Palawan is the malaria-endemic province with the highest prevalence of malaria in the Philippines, and microscopists (community health workers) have been allowed to deliver early diagnosis and prompt treatment of malaria throughout the province since 1999. To improve the quality of care, the present mixed-methods study attempted to identify the factors associated with satisfaction of patients in Palawan with their microscopists by analyzing the patients' perspectives. First, a quantitative cross-sectional study was conducted among 48 ex-patients and their nine microscopists. Ex-patients were asked about their satisfaction with care, and microscopists were asked about their job satisfaction and ability. Structural equation modeling was conducted for data analysis. Second, a qualitative cross-sectional study was performed using narrative interviews. Data were analyzed using the NVivo 10 software program. As a result, both studies revealed high patient satisfaction with microscopists. The quantitative study showed that ex-patients' satisfaction with their microscopists was independently enhanced by two factors: high ability in malaria microscopic observation and low household wealth of the patients. Particularly, specific subpopulations (those with lower household wealth, relatively old people, and ethnic minorities) were more satisfied with the microscopists' care. The qualitative study strengthens this finding by showing that their microscopists offered prompt and precise diagnosis and effective treatment for free. In conclusion, microscopists were shown to have an important role in narrowing the disparities in malaria care in Palawan. It is important to maintain/enhance the ability of microscopists in malaria microscopy to satisfy their patients.

Behind Enemy Lines: Immunomodulatory Armamentarium of the Schistosome Parasite.

The deeply rooted, intricate relationship between the Schistosoma parasite and the human host has enabled the parasite to successfully survive within the host and surreptitiously evade the host's immune attacks. The parasite has developed a variety of strategies in its immunomodulatory armamentarium to promote infection without getting harmed or killed in the battlefield of immune responses. These include the production of immunomodulatory molecules, alteration of membranes, and the promotion of granuloma formation. Schistosomiasis thus serves as a paradigm for understanding the Th2 immune responses seen in various helminthiases. This review therefore aims to summarize the immunomodulatory mechanisms of the schistosome parasites to survive inside the host. Understanding these immunomodulatory strategies not only provides information on parasite-host interactions, but also forms the basis in the development of novel drugs and vaccines against the schistosome infection, as well as various types of autoimmune and inflammatory conditions.

Field Evaluation of Recombinant Antigen ELISA in Detecting Zoonotic Schistosome Infection Among Water Buffaloes in Endemic Municipalities in the Philippines.

In this study, we investigated the use of recombinant antigens thioredoxin peroxidase-1 (rSjTPx-1) and tandem repeat rSj1TR in evaluating the antibody positivity rates of Schistosoma japonicum infection among water buffaloes from four endemic areas in the Philippines, two municipalities with high endemicity (Calatrava, Negros Occidental and Catarman, Northern Samar) and two municipalities nearing elimination with no cases of human schistosomiasis (Talibon and Trinidad, Bohol). These recombinant antigen ELISA assays were compared with other diagnostic tests including SEA-ELISA, FECT, and fecal-based PCR. Results showed that rSj1TR-ELISA has the highest agreement with PCR in all study areas. Furthermore, significant positivity rates among water buffaloes were seen in Talibon and Trinidad, indicating that water buffaloes are maintaining the schistosome parasites in transmission areas even in the absence of human infection. Hence, serological assay using a more sensitive and specific rSj1TR-ELISA can be used for animal surveillance to prevent emergence and re-emergence of human schistosomiasis.

Prevalence of Entamoeba histolytica and Entamoeba dispar among residents of an urban slum area in Manila, Philippines as detected by the polymerase chain reaction.

This study aimed to determine the prevalence of Entamoeba histolytica and Entamoeba dispar infections among residents in BASECO compound, Manila, Philippines using polymerase chain reaction (PCR). Formalin-ether concentration technique (FECT)-treated stool samples were examined under the light microscope to determine the presence of Entamoeba, helminths and other protozoan parasites. DNA was directly extracted from the FECT-treated samples and was subjected to PCR to determine E. histolytica and E. dispar infections. In this study, stool samples were collected from 2,232 residents of BASECO compound. Microscopic examination of FECT concentrated samples found 38 samples (1.703%) positive for E. histolytica/E. dispar. The E. histolytica/E. dispar microscopically positive samples were further analyzed by PCR and found 8 samples (0.358%) infected with E. histolytica and 23 samples (1.030%) infected with E. dispar. No statistically significant difference was observed in the sex distribution, while statistically significant difference was observed among the age group and area distribution of both the Entamoeba species. The results demonstrate PCR using DNA extracted from the formalin-fixed stools as an effective epidemiologic detection method of E. histolytica and E. dispar infections.

Genetic diversity and population structure of Plasmodium falciparum in the Philippines.

BACKGROUND: In the Philippines, malaria morbidity and mortality have decreased since the 1990s by effective malaria control. Several epidemiological surveys have been performed in the country, but the characteristics of the Plasmodium falciparum populations are not yet fully understood. In this study, the genetic structure of P. falciparum populations in the Philippines was examined. METHODS: Population genetic analyses based on polymorphisms of 10 microsatellite loci of the parasite were conducted on 92 isolates from three provinces (Kalinga, Palawan, and Davao del Norte) with different malaria endemicity. RESULTS: The levels of genetic diversity and the effective population sizes of P. falciparum in the Philippines were similar to those reported in the mainland of Southeast Asia or South America. In the low malaria transmission area (Kalinga), there was a low level of genetic diversity and a strong linkage disequilibrium (LD) when the single-clone haplotype (SCH) was used in the multilocus LD analysis, while in the high malaria transmission areas (Palawan and Davao del Norte), there was a high level of genetic diversity and a weak LD when SCH was used in the multilocus LD analysis. On the other hand, when the unique haplotypes were used in the multilocus LD analysis, no significant LD was observed in the Kalinga and the Palawan populations. The Kalinga and the Palawan populations were, therefore, estimated to have an epidemic population structure. The three populations were moderately differentiated from each other. CONCLUSION: In each area, the level of genetic diversity correlates with the local malaria endemicity. These findings confirm that population genetic analyses using microsatellite loci are a useful tool for evaluating malaria endemicity.

Detection of canine Schistosoma japonicum infection using recombinant thioredoxin peroxidase-1 and tandem repeat proteins.

Humans and dogs live very close together and share various pathogens causing zoonotic parasitoses like schistosomiasis. A previous population genetics study done for schistosomes in the Philippines suggested that there is a high transmission level of Schistosoma japonicum among humans and dogs proving that the latter are important reservoirs for this zoonotic parasite. A more sensitive and specific test detecting schistosome infection in dogs will therefore strengthen the zoonotic surveillance, which might help in the possible elimination of this ancient disease. In this study, recombinant thioredoxin peroxidase-1 (SjTPx-1) and tandem repeat proteins (Sj1TR, Sj2TR, Sj4TR, Sj7TR) previously tested on human and water buffalo samples were used to assess its diagnostic applicability to dogs. Fifty-nine dog serum and stool samples were collected in the schistosomiasis-endemic municipalities of Calatrava, Negros Occidental and Catarman, Northern Samar in the Philippines and examined using the ELISA as compared to microscopy and fecal sample-based PCR. Samples positive for Babesia gibsoni and Dirofilaria immitis were also used to check for cross-reaction. Results showed that SjTPx-1 (80% sensitivity, 92.3% specificity) and Sj7TR (73.3% sensitivity, 92.3% specificity) have good potentials for diagnosing S. japonicum infection in dogs. These diagnostic antigens will therefore improve the surveillance in the transmission of the parasites from dogs to humans.

Prevalence survey of schistosomiasis in Mindanao and the Visayas, The Philippines.

The first two phases of a national prevalence survey of schistosomiasis in The Philippines were completed in Mindanao in 2005 and the Visayas in the first quarter of 2007. The design was a stratified two-step systematic cluster sampling, with two Kato-Katz thick smears examined from each participant. In Mindanao, a total of 22 provinces spread in six regions were covered by the survey with five barangays (equivalent to a village) per province for a total of 110 barangays. The response rate was 70.9% with a total of 21,390 individuals examined. The province of Maguindanao, a known endemic area for schistosomiasis japonica, failed to take part in the survey. In the Visayas, 10 out of 11 provinces, spread out in three regions, participated in the survey. There were 6321 respondents for an overall participation rate of 32.2%. Mindanao showed a wider coverage of the disease than the Visayas (60% versus 45%). By region, Caraga or Region 13 ranked first in Mindanao and Region 8 in the Visayas. By province, Agusan del Sur is first on the list, followed by Northern Samar and then Eastern Samar. Overall, the prevalence rate among males is higher than that of females suggesting the occupational hazard of farming and fishing among the males. The higher exposure among farmers and fishermen is also borne out by the age distribution of the disease. Prevalence remains consistently high among the adults compared with the younger age groups. The survey also covered other helminth infections that can be detected in a stool survey, notably soil-transmitted helminthes and food-borne trematodes.

Phylogenetic relationship of Plasmodium falciparum populations in the Philippines.

Malaria is one of the major infectious diseases in the Philippines. It is being targeted for control through sustained early diagnosis, treatment and mosquito control. It is in this light that understanding the genetic background of the parasite population is important not only for basic biology of the organism but also for epidemiology and control of the disease. In the present study, molecular phylogenetic relationships of the 3 Plasmodium falciparum populations in the Philippines with the other populations in the world were inferred based on polymorphisms of 9 highly polymorphic microsatellite DNA loci in the parasite genome. A total of 92 P. falciparum isolates collected from 3 provinces (Kalinga, Palawan and Davao del Norte) in the Philippines, and 8 from other populations (3 African, 2 South American, 2 Papua New Guinean, and 1 Thai) that were previously reported, were used for the analysis. The phylogenetic tree showed that the 3 Philippine populations were genetically divergent from each other as compared to the other populations. The branching pattern of the tree suggests that the 3 Philippine populations were relatively close to the Thai population, rather than the Papua New Guinean populations, indicating that the ancestor of the 3 Philippines populations were introduced from Indochina peninsula, and not from countries located south of the Philippines such as Papua New Guinea or Indonesia.

Bottom-up approach to strengthen community-based malaria control strategy from community health workers' perceptions of their past, present, and future: a qualitative study in Palawan, Philippines.

BACKGROUND: Microscopists have active roles in bringing malaria diagnosis and treatment closer to households in Palawan, the highest malaria-endemic province in the Philippines. To accelerate the elimination of malaria in Palawan, we performed a study based on the bottom-up approach to provide profound data to strengthen this community-based malaria control from the microscopists' point of view. METHODS: We performed a qualitative cross-sectional study in Palawan. Four focus group discussions with 50 microscopists were conducted in Palawan from November 2010 to February 2011. During the discussions, the following open-ended questions were addressed: motivation for applying to be microscopists in the "Past" category; job satisfaction, role, problems, and saddest and happiest experiences working as microscopists in the "Present" category; and willingness towards task shifting in the "Future" category. Data were transcribed and analyzed by framework analysis using the NVivo software program. RESULTS: The present study innovatively proposed the following strategies: reinforcement strategy (adequate supplies and settings), highly prioritized additional strategies (improving social status of microscopists, issuing a travel budget, and including indigenous populations), regional additional strategies (additional malaria control in the southern region and task shifting in the northern region), and less prioritized additional strategies (employment policy and health checkup). CONCLUSION: A bottom-up approach using microscopists' perceptions would be a valuable method to propose practical and effective additional strategies for strengthening community-based malaria control.

Evaluation of Schistosoma japonicum thioredoxin peroxidase-1 as a potential circulating antigen target for the diagnosis of Asian schistosomiasis.

Asian schistosomiasis caused by Schistosoma japonicum is a serious zoonotic disease endemic in China, the Philippines and parts of Indonesia. Mass drug administration in endemic areas resulted to decline in disease severity and intensity. The low intensity of infection limits the use of current parasitological methods for schistosomiasis diagnosis. Detection of parasite circulating antigens might provide more informative result as it may indicate the true status of infection. In this study, S. japonicum thioredoxin peroxidase-1 (SjTPx-1) a 22 kDa secreted antioxidant enzyme expressed throughout the life stages of the parasite was evaluated for its potential use as a biomarker for schistosomiasis japonica infection. Rabbit polyclonal antibody and mouse monoclonal antibodies (mAbs) were raised against the recombinant SjTPx-1 (rSjTPx-1). The antibodies produced against the recombinant antigen was confirmed to detect the native SjTPx-1 in crude adult worm lysate. Likewise, the specific binding of mAbs to parasite TPx-1 and not to mammalian peroxiredoxin-1 orthologues was also confirmed. The double antibody sandwich ELISA developed in this study was able to detect at least 1 ng/ml of rSjTPx-1. In addition, this method was able to detect the antigen from all serum samples of experimentally infected rabbit and mice. The diagnostic potential of SjTPx-1 in human clinical samples was also evaluated, in which 4 out of 10 stool-confirmed serum samples had detectable levels of the antigen. The results suggest that SjTPx-1 can be a potential biomarker for Asian zoonotic schistosomiasis.