Treatment with insecticide and fungicide on the physiological quality of lentil seeds / Tratamento com inseticida e fungicida na qualidade fisiológica de sementes de lentilha

The low Brazilian productive index and the high demand have aroused interest in the cultivation of lentils, however the legume is little known and needs further studies. The objective of this study was to analyze and identify the effects of treatments with insecticides and fungicides on the physiological quality of lentil seeds, CA-1512 strain. The experiments were conducted in the seed laboratory in a completely randomized design with seven treatments and four replicates. Seed treatment with Thiophanate-methyl; Fluazinam® (180 ml) + Pyraclostrobin; Thiophanate-methyl; Fipronil® (150 ml) promoted higher levels of germination under accelerated aging, lower number of abnormal seedlings and longer lengths of shoot and radicle for the emergence in paper. Treatment with Carboxin; Thiram® (250 ml) + Imidacloprid® (150 ml) allowed a higher value in the first count of germination in sand, lower number of dead seeds under accelerated aging and longer root length, in the emergence in sand. Shoot length in the emergence in sand increased after seed treatment with Metalaxyl-M; Fludioxonil® (75 ml) + Pyraclostrobin; Thiophanate-methyl; Fipronil® (150 ml). Treatments with fungicides and insecticides considerably improved the physiological properties of the seeds, thus being able to guarantee greater phytosanitary qualities in the field, generating healthier seedlings and with protection against possible pests and diseases, and consequently guaranteeing greater productivity.

O baixo índice produtivo brasileiro e a alta demanda têm despertado o interesse no cultivo da lentilha, porém a leguminosa é pouco conhecida e necessita maiores estudos. Objetivou-se neste trabalho analisar e identificar tratamentos com inseticidas e fungicidas na qualidade fisiológica das sementes de lentilha, linhagem CA-1512. Os experimentos foram conduzidos no laboratório de sementes em delineamento inteiramente casualizado com sete tratamentos e quatro repetições. O tratamento de semente com Tiofanato-metílico; Fluazinam® (180 ml) + Piraclosrobina; Tiofanato-metílico; Fipronil® (150 ml) proporcionou índices mais elevados de germinação no envelhecimento acelerado, menor número de plântulas anormais e maior comprimento de parte aérea e radícula, na emergência em papel. O tratamento com Carboxina; Tiram® (250 ml) + Imidacloprido® (150 ml) possibilitou maior índice na primeira contagem de germinação em areia, menor número de sementes mortas no envelhecimento acelerado e maior comprimento de raiz, na emergência em areia. Já o comprimento da parte aérea, na emergência em areia, aumentou com o tratamento de semente Metalaxil-M; Fludioxonil® (75 ml) + Piraclosrobina; Tiofanato-metílico; Fipronil® (150 ml). Os tratamentos com fungicidas e inseticidas melhoraram consideravelmente as propriedades fisiológicas das sementes, podendo assim, garantir maiores qualidades fitossanitárias à campo, gerando plântulas mais sadias e com proteção para possíveis pragas e doenças, e consequentemente garantindo maiores produtividades.

Treatment with insecticide and fungicide on the physiological quality of lentil seeds

Abstract The low Brazilian productive index and the high demand have aroused interest in the cultivation of lentils, however the legume is little known and needs further studies. The objective of this study was to analyze and identify the effects of treatments with insecticides and fungicides on the physiological quality of lentil seeds, CA-1512 strain. The experiments were conducted in the seed laboratory in a completely randomized design with seven treatments and four replicates. Seed treatment with Thiophanate-methyl; Fluazinam® (180 ml) + Pyraclostrobin; Thiophanate-methyl; Fipronil® (150 ml) promoted higher levels of germination under accelerated aging, lower number of abnormal seedlings and longer lengths of shoot and radicle for the emergence in paper. Treatment with Carboxin; Thiram® (250 ml) + Imidacloprid® (150 ml) allowed a higher value in the first count of germination in sand, lower number of dead seeds under accelerated aging and longer root length, in the emergence in sand. Shoot length in the emergence in sand increased after seed treatment with Metalaxyl-M; Fludioxonil® (75 ml) + Pyraclostrobin; Thiophanate-methyl; Fipronil® (150 ml). Treatments with fungicides and insecticides considerably improved the physiological properties of the seeds, thus being able to guarantee greater phytosanitary qualities in the field, generating healthier seedlings and with protection against possible pests and diseases, and consequently guaranteeing greater productivity.

Resumo O baixo índice produtivo brasileiro e a alta demanda têm despertado o interesse no cultivo da lentilha, porém a leguminosa é pouco conhecida e necessita maiores estudos. Objetivou-se neste trabalho analisar e identificar tratamentos com inseticidas e fungicidas na qualidade fisiológica das sementes de lentilha, linhagem CA-1512. Os experimentos foram conduzidos no laboratório de sementes em delineamento inteiramente casualizado com sete tratamentos e quatro repetições. O tratamento de semente com Tiofanato-metílico; Fluazinam® (180 ml) + Piraclosrobina; Tiofanato-metílico; Fipronil® (150 ml) proporcionou índices mais elevados de germinação no envelhecimento acelerado, menor número de plântulas anormais e maior comprimento de parte aérea e radícula, na emergência em papel. O tratamento com Carboxina; Tiram® (250 ml) + Imidacloprido® (150 ml) possibilitou maior índice na primeira contagem de germinação em areia, menor número de sementes mortas no envelhecimento acelerado e maior comprimento de raiz, na emergência em areia. Já o comprimento da parte aérea, na emergência em areia, aumentou com o tratamento de semente Metalaxil-M; Fludioxonil® (75 ml) + Piraclosrobina; Tiofanato-metílico; Fipronil® (150 ml). Os tratamentos com fungicidas e inseticidas melhoraram consideravelmente as propriedades fisiológicas das sementes, podendo assim, garantir maiores qualidades fitossanitárias à campo, gerando plântulas mais sadias e com proteção para possíveis pragas e doenças, e consequentemente garantindo maiores produtividades.

Leguminosas como causa infrecuente de alergia alimentaria en Colombia: reporte de dos casos y revisión de la literatura / Legumes as an infrequent cause of food allergy in Colombia: report of two cases and review of the literature

La alergia alimentaria se ha venido incrementando a nivel mundial, afectando alrededor del 1,5 % a 2,5 % de los adultos y 6 % de los niños, y tiene un gran impacto en la calidad de vida de los pacientes y sus cuidadores, debido a las dietas de restricción. Los alérgenos más prevalentes son la leche, el huevo, el trigo, la soja, los frutos secos, el maní, el pescado y los mariscos. Las leguminosas mejor estudiadas son el maní y la soja; otras leguminosas como las lentejas, garbanzos y arvejas representan la quinta causa de alergia alimentaria en el área mediterránea, en Turquía y en la India, siendo menos prevalentes en otras áreas geográficas. La alergia a las leguminosas es una entidad infrecuente en Colombia, se desconoce la prevalencia en el país. Describimos los primeros dos casos de anafilaxia por lentejas reportados en el país. Ambos pacientes menores de 18 años, con reacciones adversas tras la ingesta de leguminosas, en las cuales se demuestra alergia mediada por IgE a las lentejas y además sensibilización en el primer caso a las arvejas y garbanzos, y en el segundo caso a los frijoles. Diferentes datos sobre la prevalencia se han descrito en varias áreas geográficas, siendo mayor en países con dietas mediterráneas. Las reacciones mediadas por IgE suelen aparecer incluso con el alimento altamente cocido, debido a la termo-estabilidad de las proteínas. La reactividad cruzada más frecuente se relaciona con los garbanzos y las arvejas

Food allergy has been increasing worldwide. Affects around 1.5% to 2.5% of adults and 6% of children, and has a great impact on the quality of life of patients and their caregivers, due to restricted diets. The most prevalent allergens are milk, egg, wheat, soy, tree nuts, peanuts, fish and shellfish. The best studied legumes are peanuts and soybeans; other legumes such as lentils, chickpeas and peas represent the fifth cause of food allergy in the Mediterranean area, Turkey and India, being less prevalent in other geographical areas. Allergy to legumes is not common in Colombia, the prevalence in the country is unknown. We describe the first two cases of legumes anaphylaxis reported in the country. Both patients were under 18 years of age, with adverse reactions after ingesting legumes, in which IgE-mediated allergy was demonstrated; in the first case to lentils, peas and chickpeas, and in the second case, to lentils and beans. Different data on prevalence have been described in various geographical areas, being higher in countries with Mediterranean diets. IgE-mediated reactions usually appear even with highly cooked food, due to the thermo-stability of proteins. The most frequent cross-reactivity is related to chickpeas and peas

Identification of genomic regions associated with early plant vigour in lentil (Lens culinaris)

Lentil is one of the most important food legume species, however its genetic and genomic resources remained largely uncharacterized and unexploited. In the past few years, a number of genetic maps have been constructed and marker resources have been developed in lentil. These resources could be exploited for understanding the extent and distribution of genetic variation in genus Lens and also for developing saturated and consensus genetic maps suitable for quantitative trait loci (QTL) mapping and marker-assisted selection. The present study aims to enrich polymerase chain reaction-based linkage map of F10 recombinant inbred lines (RILs) population of 94 individuals derived from cross WA8649090 9 Precoz and identification of QTLs linked to early plant vigour traits. Of the 268 polymorphic markers (93 simple sequence repeats (SSR), three inter-simple sequence repeats (ISSRs) and 172 random amplified polymorphic DNA (RAPDs)), 265 (90 SSRs, three ISSRs and 172 RAPDs) were mapped on seven linkage groups, varying in length between 25.6 and 210.3 cM, coverage of 809.4 cM with an average marker spacing of 3.05 cM. The study also reported assigning of 24 new cross-genera SSRs of Trifolium pratense on the present linkage map. The RILs along with the parents were screened for shoot length, root length, seedling length, dry weight, number of leaves and number of branches based on two replications under polyhouse conditions. A QTLhotspot consisting of six QTLs for shoot length (cm), root length (cm) and seedling length (cm) was observed between a map distances of 56.61 and 86.81 cM on LG1.

Roles of quorum sensing molecules from Rhizobium etli RT1 in bacterial motility and biofilm formation

ABSTRACT Strain RT1 was isolated from root nodules of Lens culinaris (a lentil) and characterized as Rhizobium etli (a Gram-negative soil-borne bacterium) by 16S rDNA sequencing and phylogenetic analysis. The signaling molecules produced by R. etli (RT1) were detected and identified by high-performance liquid chromatography coupled with mass spectrometry. The most abundant and biologically active N-acyl homoserine lactone molecules (3-oxo-C8-HSL and 3-OH-C14-HSL) were detected in the ethyl acetate extract of RT1. The biological role of 3-oxo-C8-HSL was evaluated in RT1. Bacterial motility and biofilm formation were affected or modified on increasing concentrations of 3-oxo-C8-HSL. Results confirmed the existence of cell communication in RT1 mediated by 3-oxo-C8-HSL, and positive correlations were found among quorum sensing, motility and biofilm formation in RT1.

Thermal processing effects on in vitro Antioxidant activities of five common Indian Pulses.

Five different types of pulses commonly used in daily cuisine in India, viz. Lentil (Lens culinaris), Green gram (Vigna radiate), Bengal gram (Cicer arietinum), Red kidney beans (Phaseolus vulgaris) and Soybean (Glycine max), were analyzed for their in vitro antioxidant profile before and after thermal processing in water. The thermal processing included methods commonly employed in India for cooking, viz. pressure cooking, microwave treatment, boiling in water and boiling after overnight soaking. The assays performed included ABTS radical decolorization assay, DPPH radical decolorization assay, hydrogen peroxide scavenging assay and hydroxyl radical scavenging assay. The antioxidant profile of the five pulses improved significantly after microwave treatment as evident from all assays except DPPH assay. In case of Bengal gram, improvement was also observed after pressure cooking. In case of soybean, however, apart from microwave treatment, no significant improvement was observed. Enhanced activity of the pulses shown after thermal processing in water might be due to enhanced extraction of polyphenols like gallo-ellagitannins, phenolic acids or flavonoid glycosides, which have less solubility in normal water but enhanced solubility in hot water. The study indicated a systematic approach to ascertain antioxidant quality of pulses after thermal treatment with water.

Comparative analysis of the antibacterial activity of some phytolectins.

The aim of this work was to analyse the comparative effects of the antibacterial properties of partially purified lectins from the seeds of Artocarpus heterophyllus (jack fruit), Canavalia ensiformis (jack bean), Lens culinaris (lentil) and Pisum sativum (pea) against the bacteria Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa. The lectins were isolated by partial purification using ammonium sulphate precipitation and dialysis. The antimicrobial activity was studied using agar well diffusion method. The results showed that the Jack fruit lectin had a potent anti-bacterial activity against S. aureus, B. subtilis, E. coli and P. aeruginosa whereas Pea and jack bean lectin were found to be effective bacteriostatic agents which reduced the growth of bacteria and lentil lectin showed the least antibacterial activity. A comparison of the antibacterial activity of phytolectins with conventional antibiotics namely ampicillin and tetracycline was also carried out. Studies revealed that the antibacterial activities of the conventional antibiotics are higher than that of the plant extracts at the same concentration in accordance to literature.

Comparative analysis of the antibacterial activity of some phytolectins.

The aim of this work was to analyse the comparative effects of the antibacterial properties of partially purified lectins from the seeds of Artocarpus heterophyllus (jack fruit), Canavalia ensiformis (jack bean), Lens culinaris (lentil) and Pisum sativum (pea) against the bacteria Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa. The lectins were isolated by partial purification using ammonium sulphate precipitation and dialysis. The antimicrobial activity was studied using agar well diffusion method. The results showed that the Jack fruit lectin had a potent anti-bacterial activity against S. aureus, B. subtilis, E. coli and P. aeruginosa whereas Pea and jack bean lectin were found to be effective bacteriostatic agents which reduced the growth of bacteria and lentil lectin showed the least antibacterial activity. A comparison of the antibacterial activity of phytolectins with conventional antibiotics namely ampicillin and tetracycline was also carried out. Studies revealed that the antibacterial activities of the conventional antibiotics are higher than that of the plant extracts at the same concentration in accordance to literature.

Pharmacognostic evaluation of Lens culinaris Medikus seeds

Objective: To present a detailed pharmacognostic study of the Lens culinaris Medikus (Fabaceae) seeds, a food grain used as Dhal in India. Methods: The macroscopy, microscopy, fluorescence analysis of powdered drug, physicochemical analysis, preliminary testing and other WHO recommended methods for standardization were investigated. Results: Seeds are greyish brown in colour. Treatment of powdered drug with various chemical reagents showed the presence of proteins, cellulose, lignins and fixed oils. Microscopy of seeds revealed the presence of starch grains in seed. The colour of seed coat changed from brown to greenish grey in day light and brown to black at 254nm when treated with acetic acid. Total ash value of the seeds was found to be 1.86% w/w whereas foreign organic matter was found to be nil. Qualitative phytochemical analysis revealed the presence of tannins and flavonoids in acetone extract. Conclusions: The present study on pharmacognostic profile of Lens culinaris Medikus seeds provides an important tool in identification and authentication of this plant to researchers in future.

Is the butterfly Tomares ballus (Lepidoptera: Lycaenidae) a potential pest of Lens culinaris (Leguminosae)?

The lentil (Lens culinaris) is identified as a new host plant of the lepidopteran Tomares ballus. Five larvae of T. ballus were found on 19 May, 2007 in a crop of "castellana" lentils in Toledo Province, Spain and reared in the laboratory. The larval brown spiracles are slightly darker than the rest of the pupa. Traditional cultural practices reduce the insect’s probability of completing its life-cycle. We present flight phenology data for T. ballus from the unedited Atlamar database (1 073 records from 1887 to 2003), based on the 438 records for which the year, month and day are known. This period coincides with the flowering and formation of the legumes of various species of leguminosae, such as lentils. The peak activity of the imagos occurs in the second half of March and the first half of April, and the last larvae must be present in the field until the beginning of June. Rev. Biol. Trop. 57 (3): 623-634. Epub 2009 September 30.

La lenteja (Lens culinaris) es una nueva planta hospedera de la mariposa Tomares ballus. Hallamos cinco larvas el 19 de mayo de 2007 en un cultivo de lentejas de la variedad "castellana" en la provincia de Toledo, España y las criamos en el laboratorio. Los espiráculos, de color marrón, son ligeramente más oscuros que el resto de la pupa. Las prácticas culturales tradicionales reducen al insecto la probabilidad de completar su ciclo de vida. Se presenta la fenología de datos de vuelo para T. ballus a partir de la base de datos Atlamar (1 073 registros de 1887 a 2003), sobre la base de los 438 registros en que se conocen año, mes y día. Este período coincide con la floración de diversas especies de leguminosas, incluyendo las lentejas. El pico de actividad de los imagos se produce en la segunda quincena de marzo y la primera quincena de abril, y las últimas larvas deben estar presentes en el campo hasta principios de junio.

Alteration of CK8 Both in Its Expression Level and Its Glycan Parts Related to HCC Metastatic Ability / 生物化学与生物物理进展

A glycomic method was used to screen the aberrantly ?1-6 fucosylated glycoproteins related to HCC metastasis and analyze the alteration of CK8 both in its expression level and its glycan parts associated with metastatic ability. Based on the approach, 2-DE coupled with lectin affinity blot, lectin affinity precipitation followed by MALDI-TOF-MS/MS, the lens culinaris agglutinin (LCA) affinity glycoprotein profiles from MHCC97-L and MHCC97-H cells, two higher metastatic HCC cell lines, were obtained, in which a differentially displayed protein spot was indicated when compared with Hep3B, in the region within 55～60 ku in molecular mass and 4～6 in isoelectric point. The identification result was CK8 by MALDI-TOF-MS/MS. To confirm the relation between increased core-fucosylation of CK8 and HCC metastasis, LCA affinity precipitation was used to extract the ?1-6 fucosylated glycoproteins, followed by Western blot. And it was found that CK8 was highly fucosylated in both MHCC97-L and MHCC97-H cells compared to Hep3B. Immunofluorescence analysis and Western blot were used to detect its intracellular localization and its protein expression levels, indicating that CK8 distributed in cytoplasm and increased protein expressions in MHCC97-L and MHCC97-H cell lines. And further lectin binding studies found that CK8 has a high affinity for Con A in both MHCC97-H and Hep3B cells, indicating that CK8 was a glycoprotein with high-mannose type N-glycans. But the amount of the lectin RCA-1 binding to CK8 was greater in MHCC97-H than Hep3B, suggesting that CK8 contained the increased terminal galactose residues ?-1, 4-linked to GlcNAc in MHCC97-H. All the results suggested that the increase of CK8 in its protein expression level, core-fucosylation and terminal gal ?1,4 GlcNAc disaccharides might be related to HCC metastatic ability.

Clinical significance of lens culinaris agglutinin-A reactive AFP for differential diagnosis of hepatocellular carcinoma and liver cirrhosis / 대한내과학회지

AFP has been the most useful tumor marker of hepatocellular carcinoma (HCC) because AFP is tumor specific and organ specific. However it can be elevated in liver cirrhosis (LC) and chronic hepatitis as well. AFP in hepatocellular carcinoma has been known to be highly reactive Lens Culinaris Agglutinin-A (LCA-A). The aim of our study is to find whether LCA-A reactive AFP (AFP-L3) is useful in differentiation of HCC and LC with high serum AFP level (>20 ng/ml). METHODS: The material consists of sera from 18 patients with HCC and 16 patients with LC whose serum AFP concentration was above 20 ng/ml. Sera were tested for AFP-L3 by lectin-affinity electrophoresis, coupled with antibody affinity blotting with alpha-fetoprotein differentiation kit L (Wako Pure Chemical Industries, Osaka, Japan). AFP-L3 fraction percentage was checked by densitometry (580nm, Cliniscan II, Helena). RESULTS: AFP-L3 detection rate in membrane was 61.1% in HCC and 37.5% in LC. When cut-off value level is 15%, the positive rate was 61.1% in HCC and 31.2% in LC, and sensitivity and specificity of AFP-L3 fraction in HCC was 61.1% and 68.8% respectively. In according to these results, AFP-L3 was a useful marker in differentiation HCC and LC with high serum AFP level. There was no significant relation in AFP-L3 fraction and tumor size, but there was significant relation between AFP-L3 fraction and total AFP serum level(P=0.049). CONCLUSION: These results lead us to conclude that the level of AFP-L3 is a useful marker in differentiation of HCC and LC with high serum AFP level.

Clinical significance of lens culinaris agglutinin-A reactive AFP for differential diagnosis of hepatocellular carcinoma and liver cirrhosis / 대한내과학회지

AFP has been the most useful tumor marker of hepatocellular carcinoma (HCC) because AFP is tumor specific and organ specific. However it can be elevated in liver cirrhosis (LC) and chronic hepatitis as well. AFP in hepatocellular carcinoma has been known to be highly reactive Lens Culinaris Agglutinin-A (LCA-A). The aim of our study is to find whether LCA-A reactive AFP (AFP-L3) is useful in differentiation of HCC and LC with high serum AFP level (>20 ng/ml). METHODS: The material consists of sera from 18 patients with HCC and 16 patients with LC whose serum AFP concentration was above 20 ng/ml. Sera were tested for AFP-L3 by lectin-affinity electrophoresis, coupled with antibody affinity blotting with alpha-fetoprotein differentiation kit L (Wako Pure Chemical Industries, Osaka, Japan). AFP-L3 fraction percentage was checked by densitometry (580nm, Cliniscan II, Helena). RESULTS: AFP-L3 detection rate in membrane was 61.1% in HCC and 37.5% in LC. When cut-off value level is 15%, the positive rate was 61.1% in HCC and 31.2% in LC, and sensitivity and specificity of AFP-L3 fraction in HCC was 61.1% and 68.8% respectively. In according to these results, AFP-L3 was a useful marker in differentiation HCC and LC with high serum AFP level. There was no significant relation in AFP-L3 fraction and tumor size, but there was significant relation between AFP-L3 fraction and total AFP serum level(P=0.049). CONCLUSION: These results lead us to conclude that the level of AFP-L3 is a useful marker in differentiation of HCC and LC with high serum AFP level.

Usefulness of Lens Culinaris Agglutinin-A Reactive Alpha-Fetoprotein for the Diagnosis of Hepatocelluar Carcinoma in Patients with Chronic Liver Disease Who Showed Significant Increment of Serum Alpha-Fetoprotein Level and No Mass Lesion in the Liver on U / 대한간학회지

BACKGROUND/AIMS: Lens culinaris agglutinin-A reactive alpha-fetoprotein (AFP L3) has been reported to be highly specific for the diagnosis of hepatocellular carcinoma (HCC). The present study was to evaluate the clinical usefulness of AFP-L3 for the diagnosis of HCC in the patients either with chronic liver disease or with HCC in complete remission who showed significant increment of serum AFP level and no mass lesion in the liver on ultrasonography. METHODS: A total numer of 34 patients (24 with chronic liver disease, 10 with HCC in complete remission) were enrolled, who showed significant increment of serum AFP level and no mass lesion in the liver on ultrasonography. Serum AFP L3 levels were analysed by AFP differentiation kit L. Abdominal spiral CT or ultrasonogram was performed at 1-3 month intervals and all of the patients were followed up for more than 6 months. RESULTS: Among 24 patients with chronic liver disease, two were positive (higher than 15%) for AFP L3; however, HCC was not detected in these patients, while HCCs were detected in two of 22 patients negative for AFP L3 during followe-up. Eight of the 10 patients with HCC in complete remission were positive for AFP L3; recurrent HCCs were detected in 7 of those 8 patients as well as in the rest 2 patients negative for AFP L3 during follow-up. The overall sensitivity and specificity of AFP L3 measurement for the detection of HCC within 6 months of follow-up were 63.6% and 87.0%, respectively. The positive and negative predictive value for HCC in patients with chronic liver disease were O% and 90.9% and for recurrent HCC in HCC patients in remission were 87.5% and ON, respectively. CONCLUSION: The measurement of AFP L3 is suggested to be useful for the diagnostic strategy in patients either with chronic liver disease or hepatocellular carcinoma in complete remission, who showed progressive increment of serum AFP level and no mass lesion in liver on ultrasonogram.

The Preparation and Application of Anti-human Heterogeneous Lens culinaris Agglutinin Reactive AFP Monoclonal Antibodies / 第二军医大学学报

In this paper, we report the study concerning the preparation and application of anti-human heterogeneous AFP-R-LCA monoclonal antibodies (VG5 VD12 VB5 VA8 VD12). These McAbs had higher affinity and specificity to AFP-R-LCA than the routinely used anti-AFP polyclonal antibody. Competitive assays indicated that they could bind to new antigenic determinants which are different from the previously reported a and b sites. In an assay of the McAbs, we first set up a two site sandwich ELISA method and then tested, using this method, serum samples from 69 patients with PHC, 67 patients with benign liver diseases, 30 pregnant women and 30 normal controls. The results showed that this two site sandwich ELISA method had low pseudo-positive rate. The detection range was 5 1000 ng/ml, with the properties of simplicity, accuracy and reproducibility. It could be used not only for the early diagnosis of PHC in patients with AFP levels less than 400 ng/ml, but also for the differential diagnosis of PHC from benign liver diseases.