旋毛虫成囊前期幼虫抗原保护性免疫的研究

目的比较旋毛虫成囊前期幼虫虫体抗原、排泄分泌抗原和表面抗原对小鼠产生的免疫保护作用。方法分别用旋毛虫成囊前期幼虫虫体抗原、排泄分泌抗原、表面抗原免疫小鼠,同时设佐剂组和阴性对照组,间隔7d免疫1次,共3次。末次免疫后7d,每只小鼠用200条旋毛虫感染期幼虫经口进行攻击感染。感染后7d和30d分别检查各组小鼠肠道成虫数和肌幼虫数;用ELISA测血清中抗旋毛虫肌幼虫IgG抗体。结果虫体抗原、排泄分泌抗原、表面抗原和佐剂组的成虫减虫率分别为84．89％、89．73％、85．65％、2．57％;肌幼虫减虫率分别为71．71％、80．98％、73．66％、5．60％。排泄分泌抗原组、表面抗原组的成虫减虫率（P均〈0．05）及肌幼虫减虫率（P均〈0．01）均高于虫体抗原组。各免疫组小鼠血清IgG抗体滴度明显升高,虫体抗原组、排泄分泌抗原组和表面抗原组的几何平均倒数滴度分别为2798．89、3474．51、2984．83,分别为阴性对照组（459．32）的6．09、7．56、6．50倍。结论旋毛虫成囊前期幼虫虫体抗原、排泄分泌抗原和表面抗原均能诱导宿主产生较强的抗攻击感染保护力。成囊前期幼虫的排泄分泌抗原显示出更强的免疫原性。     

The distribution of ABO system blood groups in patients with unilocular echinococcosis

The distribution of the AB0 blood groups was investigated in 624 patients with echinococcosis operated on and in 579 persons whose sera were subjected to IHAT with echinococcosis diagnosticum. Higher disease incidence was recorded in patients with the A (II) blood, especially in males, as compared to the controls. Simultaneous involvement of the liver, lungs and other organs was more common in them. Rh-positive patients predominated both in the A (II) and control groups. Blood group distribution with regard to IHAT findings failed to reveal any regular features.     

旋毛虫排泄分泌抗原对小鼠的保护性免疫

目的比较旋毛虫成虫排泄分泌抗原(ES抗原)、肌幼虫ES抗原、成虫和肌幼虫ES混合抗原对小鼠的免疫保护作用。方法用生理盐水培养法从培养液中提取成虫ES抗原、肌幼虫ES抗原,分别用成虫ES抗原、肌幼虫ES抗原、成虫和肌幼虫ES混合抗原免疫小鼠,同时设佐剂组和对照组,间隔7d共免疫3次。末次免疫后7天,每只小鼠用200条旋毛虫感染期幼虫经口进行攻击感染。感染后7天和30天检查各组小鼠肠道成虫数和肌幼虫数。结果旋毛虫成虫ES抗原组、肌幼虫ES抗原组、成虫和肌幼虫ES混合抗原组的成虫减虫率分别为87.95%、69.48%、84.34%,肌幼虫减虫率分别为74.79%、87.97%、86.87%。成虫ES抗原组、成虫与肌幼虫ES抗原混合组的成虫减虫率均高于肌幼虫ES抗原组(P均<0.05)。肌幼虫ES抗原组、成虫与肌幼虫ES抗原混合组的肌幼虫减虫率均高于成虫ES抗原组(P均<0.01)。结论旋毛虫成虫和肌幼虫ES混合抗原均能诱导小鼠产生抗成虫及肌幼虫较强的免疫力。     

旋毛虫排泄分泌抗原的成分及应用价值

旋毛虫病是一种重要的人畜共患寄生虫病，长期以来用于免疫诊断和免疫预防的旋毛虫抗原，尤其是排泄分泌（ES）抗原引起了国内外众多学者的关注。本文综述了近年来旋毛虫ES抗原有效成分的研究及其在免疫诊断和免疫预防方面的应用价值，并且提出应用DNA重组技术在体外大量表达ES抗原和利用多肽合成技术在体外大量合成抗原用于免疫诊断和免疫预防将是今后的研究方向。     

旋毛虫成虫抗原的免疫保护性研究

目的 比较旋毛虫成虫虫体抗原、排泄分泌抗原和表面抗原对小鼠产生的免疫保护作用。 方法 检查免疫鼠和对照鼠肠道成虫、肌幼虫和血液中的嗜酸性粒细胞数;用ELISA测血清中抗旋毛虫肌幼虫IgG抗体滴度。 结果旋毛虫成虫虫体抗原、排泄分泌抗原和表面抗原免疫组的成虫减虫率分别为84.48％、89.98％和85.16％;肌幼虫减虫率分别为69.82％、78.80％和73.94％。3种抗原免疫组小鼠血中的嗜酸性粒细胞(EOS)数明显增多,血清中IgG抗体滴度明显升高,IgG抗体的几何平均倒数滴度(GMRT)分别是未免疫组的6.96、7.99和6.06倍。 结论 旋毛虫成虫虫体抗原、排泄分泌抗原和表面抗原均能诱导宿主产生较强的抗攻击感染保护力,且可激发特异性体液免疫和细胞免疫。成虫的排泄分泌抗原显示出更强的免疫原性。     

Seroprevalence of Trichinella in slaughter pigs in Kathmandu Valley, Nepal

The aim of the present study was to determine the Trichinella seroprevalence in slaughter pigs in Kathmandu Valley, Nepal. Serum samples were obtained from 400 pigs at 4 major slaughterhouses and tested for Trichinella antibodies by ELISA using larval excretory-secretory (E/S) antigen. Four were positive and one was equivocal, giving a Trichinella seroprevalence of 1% (95% CI: 0.27 - 2.54). On titration, all positive and equivocal samples had titers greater than 1:80. Upon re-examination the equivocal sample failed to give a positive ELISA result. The pigs were from four major areas of Nepal, Kathmandu Valley, eastern Nepal, Terai and adjoining areas of the valley. Positive results were found from only Kathmandu Valley and adjoining areas. There was no significant difference in the prevalence between areas (p = 0.43). All four positive samples were from indoor managed pigs. The Trichinella seroprevalence determined in this study deserves a direct demonstration of the parasites for proof of the presence of Trichinella in Nepal and to discover the species and infection sources.     

旋毛虫成囊前期幼虫排泄分泌抗原的研究进展

在旋毛虫感染过程中,成囊前期幼虫（PEL）是旋毛虫侵入宿主肌肉的侵入期。旋毛虫PEL比成囊期幼虫（EL）在宿主体内约早2周出现。成囊前期幼虫抗原（PELA）对旋毛虫病的早期免疫学诊断具有较高的敏感性。而旋毛虫排泄分泌抗原具双重免疫功能,即有良好的抗原性和免疫原性。因此,本文就旋毛虫成囊前期幼虫ES抗原研究进展做一综述。     

The usefulness of ELISA for diagnosis of trichinellosis in pigs and wild boars

ELISA can be used to measure produced antibodies or Trichinella spp. antigens in the samples. They are detected with antibodies linked to an enzyme that reacts with a substrate and generate a colour reaction. The optical density (OD) of the reaction is measured spectrophotometrically. ELISA assays can be done in several different procedures called "direct", "indirect", "sa ndwich", and "competition" ELISA. Since the 1970s, the studies have been done on improving or replacing direct methods of Trichinella diagnosis with serological methods based on the ELISA. When somatic antigens of L1 T. spiralis were used, the specificity of the ELISA was poor due to a high probability of cross-reactions with other pathogens. During the 1980s the specificity of the ELISA was improved by excretory-secretory (E/S) antigens obtained during Trichinella muscle larvae incubation in vitro. Recently a synthetic glycan antigen has been developed and the increasing of ELISA specificity and sensitivity was noticed. The sensitivity of the ELISA using an E/S antigen ranging from 93.1 to 99.2% but the specificity from 90.6 to 99.4%. The ELISA method is relatively simple to apply, reliable, readily standardized and provides an acceptable balance of sensitivity and specificity. But all modified procedures should be validated. In Poland, the studies on the usefulness of ELISA for antibodies detection against T. spiralis in pigs and wild animals are limited. Own ELISA procedure was prepared in Pathophysiology Lab. in W. Stefański Institute of Parasitology of PAS. ELISA was used to examine IgG level against L1 T. spiralis in pigs and wild boars serum samples. Of 1474 pig samples, only 12 were positive. Of 1880 wild boar samples only 14 were positive. The results of this study are comparable with performance obtained using commercial sets. The results showed the usefulness of ELISA for T. spiralis diagnosis in pigs and wild boars and confirmed the possibility of use the ELISA test for application in the slaughterhouse.     

以重组蛋白为抗原的ELISA法检测旋毛虫抗体

目的 寻求特异性强、敏感性高的旋毛虫病诊断抗原。 方法 以旋毛虫新生幼虫期特异性T668基因在E.coli高效表达的重组蛋白为抗原，分别以兔、猪和健康者血清及旋毛虫病阳性血清为一抗，以辣根过氧化物酶（HRP）标记的山羊抗兔IgG、山羊抗猪IgG和山羊抗人IgG为二抗，建立检测旋毛虫抗体的间接ELISA方法，并以旋毛虫肌幼虫排泄?鄄分泌（ES）抗原作为检测对照。 结果 以T668重组蛋白为抗原，对兔、猪和人旋毛虫病血清进行检测，阳性检出率为100 ％，且敏感性高（0.016 μg / 孔），与ES抗原检测结果完全一致。 结论 T668重组抗原有望替代ES抗原检测旋毛虫抗体。     

旋毛虫感染兔唾液中抗旋毛虫IgG抗体水平

将28只日本大耳兔随机分为实验组(20只)和对照组(8只),实验组用旋毛虫脱囊幼虫经口灌胃日本大耳兔(3000条/只),对照组不做任何处理。采集感染前和感染后1～6周兔唾液和血清以及对照组兔唾液和血清。建立旋毛虫肌肉幼虫排泄分泌抗原(MLESA)为诊断抗原的间接ELISA,测定兔唾液和血清中抗旋毛虫IgG抗体。结果显示,感染后1～6周,唾液阳性率分别为10%、15%、40%、65%、85%和95%;血清阳性率分别为35%、50%、80%、90%、100%和100%。感染后1～3周,唾液阳性率与血清阳性率差异有统计学意义(χ2=3.58、5.23、6.67,P0.05),感染后4～6周,两者差异无统计学差异(χ2=0.12、1.03、1.03,P0.05)。提示在血清标本采集困难的情况下,MLESA的间接ELISA法检测唾液中抗旋毛虫IgG抗体可作为旋毛虫病免疫诊断的辅助方法。     

旋毛虫成虫可溶性抗原和排泄分泌抗原对小鼠免疫保护作用的比较研究

目的　比较旋毛虫成虫可溶性抗原和排泄分泌抗原对小鼠的免疫保护作用。方法　收集人工感染大鼠小肠内的成虫, 经研磨和冻融制备成虫可溶性抗原。采用体外培养的方法从培养液中提取旋毛虫成虫排泄分泌抗原。分别用两种抗原免疫小鼠, 间隔１ 周共免疫３ 次, 末次免疫后１ 周, 每只小鼠攻击感染１００ 条旋毛虫感染性肌肉幼虫。感染后１ 周检查小鼠小肠内成虫数量和雌虫生殖力, 感染后５ 周检查肌肉幼虫负荷。结果　成虫可溶性抗原诱导的成虫减虫率、新生幼虫减虫率和肌肉幼虫减虫率分别为７９５５ ％ 、６２２５ ％ 和６５０ ％ 。成虫排泄分泌抗原诱导的成虫减虫率、新生幼虫减虫率和肌肉幼虫减虫率分别是９７２７ ％ 、８６６０ ％ 和９００ ％ 。结论　实验结果表明旋毛虫成虫可溶性抗原和成虫排泄分泌抗原均能够诱导宿主产生较强的抗攻击感染的免疫力, 但后者的免疫原性更强。     

原头节抗原IFAT和IEST诊断人体包虫病的比较研究

以冰冻保存的绵羊棘球蚴内原头节为抗原作间接荧光抗体试验(IFAT)和间接免疫酶染色试验(IEST),检测对包虫病人、非包虫病人及键康人的敏感性、特异性及交叉反应性,并与间接血凝试验相比较(IHAT)。结果表明IFAT的敏感性和特异性分别为90.36％和97.67％;IEST的敏感性和特异性分别为96.55％和100.0％;两种方法均高于IHAT的敏感性(85.54％)和特异性(95.35％)。三种方法对血吸虫感染者和癌症病人血清均无交叉反应,但对囊虫病人的交叉反应率为IFAT(51.35％)>IHAT(37.84％)>IEST(21.62％)。以原头节为抗原IEST诊断人体包虫病敏感性高、特异性强,适用范围广,不需特殊设备,具有现场应用价值。     

The estimation of different ELISA procedures for serodiagnosis of human trichinellosis

INTRODUCTION: The most important confirmative diagnostic test for trichinellosis is the presence of the muscle larvae in a tissue biopsy but this direct method has a low sensitivity of light and moderate infections. The aim of presented study was to compare the usefulness of the results obtained by three ELISA procedures for Trichinella spp. diagnosis in human outbreaks. MATERIALS AND METHODS: All sera (cases and controls) were tested for anti-Trichinella antibodies (immunoglobulin G) using commercially available Novatec KIT and two other ELISA procedures based on excretory-secretory (ES) antigens on Trichinella spiralis muscle larvae. The main differences in ELISA procedures were: the protein concentration in antigen, dilution of human serum samples, conjugate and the time of conjugate incubation. Additional differences were noticed in ES antigen preparation procedures as well as in T. spiralis isolates used in these procedures. Serum samples were obtained from 22 symptomatical patients from Poznafi region (West Poland), geographic area where human outbreak had occurred. Control serum samples were obtained from 20 patients from an open population from a non endemic trichinellosis area. RESULTS: The results were analyzed in terms of both: statistical and epidemiological point of view. Linear regression analysis and correlations coefficient r between OD values of total 22 patients obtained in three ELISA procedures were positive and high statistically significant. Three ELISA procedures revealed different cut-off values and positivity rates for outbreak. However, the majority of positive samples were found as positive in three procedures, but some of them were positive in two or one procedure only. These individual variability in sera reactivity observed in three ELISA procedures could be very important from epidemiological point of view.     

滤纸干血滴间接荧光抗体试验应用于人体包虫病诊断和流行病学调查的研究

1986～1988年我们应用绵羊棘球蚴囊壁冰冻切片抗原作间接荧光抗体试验分别检测了13例包虫病人血清和滤纸干血滴,结果均为阳性,阳性率为100％(13/13);24例健康人血清和滤纸干血滴的阴性率分别为95.8％(23/24)和100％(24/24)。本方法敏感性和特异性较高,可节省抗原材料,采血简便易行,是包虫病较为理想的辅助诊断方法。为了研究其实用价值,我们又检测了甘肃省天祝县251人,漳县242人和会宁县683人,阳性率分别为4.78％(12/251),5.37％(13/242)和1.17％(8/683),其结果与间接血凝试验的结果(阳性率分别为4.78％,4.96％和0.9%)之间无显著性差异(P>0.05)。作者认为将该方法用于包虫病流行区人群的血清流行病学调查,可反映人群包虫病的感染率。     

The measurement of antigens released by radiation-attenuated Trichinella spiralis larvae

Summary A radioimmunoassay has been developed that uses antisera raised to different excretory-secretory antigens of infective larvae of Trichinella spiralis (LESA) to measure accurately the output of these antigens following gamma irradiation at doses from 10 to 120 Krads. In the lower range (up to 20 Krads) irradiation results in the increased export of antigens to the culture supernatant in a subsequent 3 h period, without obvious or gross damage to the worms. Higher doses (> 40 Krads) suppress antigen release over the same period compared with the activity of untreated (control) cultures. This work makes two contributions. It describes a sensitive assay system which detects and measures parasite antigens that may be important both in protection and in serodiagnosis, and it offers for the first time an explanation for the special properties of the lower dose range larval irradiation-attenuated vaccine in inducing a high degree of reinfection resistance, as reported in older literature and recently confirmed by us.     

Detection of circulating parasite antigens in murine gnathostomiasis by a two-site enzyme-linked immunosorbent assay.

A two-site enzyme-linked immunosorbent assay (ELISA) was developed for the detection of Gnathostoma spinigerum antigens in the sera of parasitized mice. This assay used IgG fractions prepared from serum of a G. spinigerum-infected rabbit as the capture antibody. The same IgG fractions were labeled with alkaline phosphatase and used as an antibody probe. The antigen detection assay was performed along with an antibody detection assay during the course of G. spinigerum infection in mice. Circulating antigen was detected after the first week of infection. The amount of detectable antigen increased steadily until the fourth week, but no significant amount of circulating antigen was detected thereafter. Serum antibody first appeared at the second week. Its level increased steadily until the fourth week, then remained high for at least eight weeks. The sensitivity of the two-site ELISA was approximately 6.75 ng/ml of larval somatic antigen and 27 ng/ml of excretory-secretory antigen. The assay gave false-positive results with Opisthorchis, Trichinella, and Angiostrongylus antigens at the level of 1, 728, 432, and 864 ng/ml or higher, respectively. This antigen detection assay may have application in the diagnosis of human gnathostomiasis.     

Comparison of three antigen preparations to detect Trichinellosis in live swine using IgG-ELISA

A swine infected with Trichinella spiralis is a source of transmission to human through consumption of raw or improperly cooked pork. Detection of larvae is suitable for carcasses, so that pigs in households or farms can be examined serologically for trichinellosis. This study compared antigens, crude (CAg), excretory-secretory (ESAg) and surface (SAg), for their potential use in IgG-ELISA. Serum samples were collected from 5 experimentally infected swine with T. spiralis (pTs), 147 positive cases of 9 other parasitic infections, 12 mixed infections of other parasites, and 35 normal controls. At the same 100% sensitivity, specificity of tests was in a range of 98-77%. ESAg was the best source of antigen with specificity of 98.3% at cut-off value of 0.439. False positives included coccidiasis (1/86) and mixed infections (2/39). For CAg, trichuriasis (2/11), coccidiasis (5/86), and mixed infections (8/39) gave cross-reactions and some of these samples had OD values far above cut-off value of 0.332. Cross-reactions of SAg were Oesophagostomum spp-like GI-nematode infection (1/1), unidentified GI-nematode infections (2/3), trichuriasis (5/11), coccidiasis (29/86) and mixed infections (4/39). Thus, ESAg has the highest potential in serodiagnosis, with antibody to T. spiralis in pigs being detected at the earliest 16 day post-infection. However, crude antigen demonstrated a good specificity at 91.8%, and this antigen has a potential to be used as a detection of choice for swine trichinellosis, but the antigen preparation must be improved for higher specificity.     

免疫血清对旋毛虫感染性幼虫侵入肠上皮细胞及其发育的影响

目的观察旋毛虫感染性幼虫排泄分泌(ES)抗原与旋毛虫感染性幼虫表面抗原免疫鼠血清对幼虫侵入HCT-8肠上皮细胞及其发育的影响。方法将旋毛虫感染性幼虫接种至半固体培养基(RPMI1640培养基+1.75%琼脂糖)+HCT-8细胞中,37℃5%CO2培养12、24、36、72和96h后镜下观察幼虫发育情况;将幼虫接种至含免疫血清的半固体培养基+HCT-8细胞中,培养15min后镜下观察幼虫形态及其对肠上皮细胞的侵入情况,36h后应用间接荧光抗体试验(IFAT)观察幼虫蜕皮,并计数Ⅰ期和Ⅱ～Ⅳ期幼虫。结果幼虫在半固体培养基培养12h可侵入HCT-8细胞单层,36～72h幼虫可蜕皮1～2次,培养96h可见早期成虫。在含ES抗原免疫血清与感染鼠血清条件下培养15min,幼虫头端可见免疫复合物形成的帽样结构,但在含表面抗原免疫血清、正常鼠血清或不含免疫血清条件下培养的幼虫头端则无帽样结构,头端带有帽样结构的幼虫不能侵入HCT-8细胞单层。在含ES抗原与表面抗原免疫血清条件下发育至Ⅱ～Ⅳ期幼虫的百分比(2.25%、2.2%)均明显低于含正常鼠血清条件下培养的幼虫(24.7%)(P0.05)。结论旋毛虫ES抗原免疫血清可阻止幼虫对肠上皮细胞的侵入,ES抗原及表面抗原免疫血清均可阻止部分幼虫的发育(蜕皮)。     

Comparative immunochemical analysis of a complete somatic extract, somatic fractionated and excretory-secretory proteins and antigens of T. spiralis larvae

The paper deals with the analysis of the protein composition of invasion Trichinella larvae, the immunochemical characteristics ofTrichinella antigens of varying molecular mass, the development of the optimum procedures for having the preparative quantities of immunogenic, the so-called major Trichinella antigens that are the major component for the production of diagnostic and prophylactic agents. Twenty nine protein bands were detected in the composite of a complete somatic Trichinella extract. It was ascertained that the most immunogenic peptides with molecular masses of 30, 43, 52-57, and 63-69 kDa were present in all Trichinella protein products while peptides with molecular masses of 43, 50-55 kDa are major antigens both in the fractionated somatic and excretory-secretory proteins in all incubation periods.     

The usefulness of ELISA test for early serological detection of Trichinella spp. infection in pigs

Trichinellosis is a parasitic zoonosis transmitted to humans through consumption of raw or undercooked meat from animals infected with nematodes of the Trichinella genus. Every year seropositive cases are found among the human population and thus trichinellosis still remains an epidemiologically important disease. The first step of the study was the optimization of a new ELISA method enabling an early and specific serological diagnosis of trichinellosis in pigs and wild boars using excretory-secretory (ES) antigens obtained from in vitro cultures of L1 T. spiralis. Serum samples were assayed for anti-T. spiralis IgG antibodies using the new ELISA protocol and a reference test--Standard manufactured by Institut Pourquier. The optimization involved the selection of suitable plates for antigen coating, dilution of sera and antibodies and their time of incubation. On the basis of the optimization a new ELISA procedure for the detection of IgG and IgM against T. spiralis was elaborated. Conventional, Iberian pigs and SPF (Specific Pathogen Free) pigs were infected with 200, 1000 and 20,000 muscle larvae of T. spiralis. Serum samples were obtained at 5 and 1 dbi (day before infection), and 5, 10, 15, 20, 25, 30, 40, 50, 60 dpi (day post infection) and screened for specific IgG antibodies against excretory-secretory L1 T. spiralis antigens. Serum samples were obtained from the EU project Trichiporse: "Safe pork and horse meat on EU markets: early and unbiased diagnostic tests for Trichinella". Field samples of conventional pigs (1474) and wild boars (1784) were obtained from slaughter houses in different parts of Poland. Pigs were examined for the presence of Trichinella spp. using the artificial digestion method. Only four pigs were naturally infected with T. spiralis, the remaining were Trichinella larvae free. ELISA was used to examine IgG levels against L1 T. spiralis in pig and wild boar sera. The usefulness of ELISA for anti-IgG detection in pigs is usually limited by the nature of the antigen. The antigens were prepared in different laboratories: in Germany--Ag ES L1 T. spiralis (N), Italy--Ag ES L1 T. spiralis (W)) and in Poland--Ag ES L1 T. spiralis. Cut-off values for ELISA along with the estimated sensitivity and specificity were calculated using different methods: S/P%, M+3SD and ROC (Receiver Operating Characteristic). In SPF and Iberian pigs inoculated with 200, 1000 and 20,000 L1 T. spiralis, specific antibodies were detected 40, 30 and 25 dpi, respectively, with the use of the Standard (reference test). The analysis of the two ELISA procedures demonstrated a high sensitivity and specificity for the newly elaborated test utilizing the Ag ES L1 T. spiralis. In conventional pigs infected with 20,000 L1 T. spiralis specific antibodies were detected from 20 dpi when employing the new protocol. Similar results for the Standard and new ELISA test were obtained for serum samples of conventional pigs infected with 200 and 1000 larvae, which became positive from 40 dpi and 30 dpi, respectively. The results showed that both: the Standard and new protocols were comparable, and based on this, the new test was applied for further research. Results obtained adopting the new protocol with three antigens showed that two of them: Ag ES L1 T. spiralis (W) and Ag ES L1 T. spiralis are similar. The specific IgG antibodies for infective doses of 200 and 1000 larvae for these antigens were detectable 40 and 30 dpi respectively. In pigs infected with the highest dose of T. spiralis larvae IgG antibodies were detectable from 20 dpi when Ag ES L1 T. spiralis was used. These results strongly indicate that in examined pigs, the specific IgG response to T. spiralis infection is dose dependant. Of 1474 examined pig sera only 0.99% gave a positive signal against ES L1 T. spiralis antigen. Of 1784 examined wild boars sera only 0.68 % gave positive results using the new ELISA protocol. ELISA is a useful method for detecting specific IgG antibodies in pigs experimentally infected with different doses of T. spiralis and naturally infected pigs. In pigs the specific IgG response is dose dependant. The Ag ES L1 T. spiralis increases the specifity of the method and reduces false positive results. Simultaneous use of both methods: digestion and ELISA for the diagnosis of Trichinella in naturally infected pigs and wild boars may increase the chances of eliminating meat infected with T. spiralis larvae.