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Characterization of a Trichinella spiralis putative serine protease. Study of its potential as sero-diagnostic tool
- Source :
- PLoS Neglected Tropical Diseases, Vol 12, Iss 5, p e0006485 (2018), PLoS Neglected Tropical Diseases
- Publication Year :
- 2018
- Publisher :
- Public Library of Science (PLoS), 2018.
-
Abstract
- Background Trichinellosis is a serious zoonositc parasitosis worldwide. Because its clinical manifestations aren’t specific, the diagnosis of trichinellosis is not easy to be made. Trichinella spiralis muscle larva (ML) excretory–secretory (ES) antigens are the most widely applied diagnostic antigens for human trichinellosis, but the major drawback of the ES antigens for assaying anti-Trichinella antibodies is the false negative in the early Trichinella infection period. The aim of this study was to characterize the T. spiralis putative serine protease (TsSP) and to investigate its potential use for diagnosis of trichinellosis. Methodology/Principal findings The full-length TsSP sequence was cloned and expressed, and recombinant TsSP (rTsSP) was purified by Ni-NTA-Sefinose Column. On Western blotting analysis the rTsSP was recognized by T. spiralis-infected mouse serum, and the natural TsSP was identified in T. spiralis ML crude and ES antigens by using anti-rTsSP serum. Expression of TsSP was detected at various T. spiralis developmental stages (newborn larvae, muscle larvae, intestinal infective larvae and adult worms). Immunolocalization identified the TsSP principally in cuticles and stichosomes of the nematode. The sensitivity of rTsSP-ELISA and ES-ELISA was 98.11% (52/53) and 88.68% (47/53) respectively (P > 0.05) when the sera from trichinellosis patients were examined. However, while twenty-one serum samples of trichinellosis patients’ sera at 19 days post-infection (dpi) were tested, the sensitivity (95.24%) of rTsSP-ELISA was distinctly higher than 71.43% of ES-ELISA (P < 0.05). The specificity (99.53%) of rTsSP-ELISA was remarkably higher than 91.98% of ES-ELISA (P < 0.01). Only one out of 20 serum samples of cysticercosis patients cross-reacted with the rTsSP. Specific anti-Trichinella IgG in infected mice was first detected by rTsSP-ELISA as soon as 7 dpi and antibody positive rate reached 100% on 10 dpi, whereas the ES-ELISA did not permit detection of 100% of infected mice before 16 dpi. Conclusions The rTsSP is a potential early diagnostic antigen for human trichinellosis.<br />Author summary Trichinellosis is an important parasitic zoonosis, and has a public health hazard and an economic impact on the safety of animal food. The diagnosis of trichinellosis is difficult and it is often misdiagnosed. There is an evident 2–3 week window stage between clinical manifestations and the anti-Trichinella IgG positive. Serine protease is a superfamily of proteolytic enzymes and exerts a major role in tissue invasion, larval development and survival of the parasites. A T. spiralis putative serine protease (TsSP) was characterized in ES proteins of T. spiralis intestinal infective larvae and adult worms by the immunoproteomics with early infection serum. In this study, the TsSP was expressed and purified. The results revealed that the TsSP was expressed at various T. spiralis stages (newborn larvae, muscle larvae, intestinal infective larvae and adult worms) and it was principally located in cuticle and stichosome of the nematode. The rTsSP was sensitive and specific for detection of anti-Trichinella IgG, and could be regarded as an early diagnostic marker of trichinellosis.
- Subjects :
- 0301 basic medicine
Life Cycles
Nematoda
Physiology
Trichinella
Helminth genetics
Biochemistry
Mice
Blood serum
Larvae
Immune Physiology
Medicine and Health Sciences
Enzyme-Linked Immunoassays
Nematode Infections
Mice, Inbred BALB C
Immune System Proteins
biology
lcsh:Public aspects of medicine
Eukaryota
Trichinellosis
Helminth Proteins
Proteases
Enzymes
Infectious Diseases
Female
Antibody
Research Article
lcsh:Arctic medicine. Tropical medicine
lcsh:RC955-962
Trichinella spiralis
Immunology
Antibodies, Helminth
Enzyme-Linked Immunosorbent Assay
Trichinosis
Cross Reactions
Research and Analysis Methods
Antibodies
Microbiology
03 medical and health sciences
Antigen
parasitic diseases
medicine
Parasitic Diseases
Helminths
Animals
Humans
Immunoassays
Public Health, Environmental and Occupational Health
Organisms
Biology and Life Sciences
Proteins
lcsh:RA1-1270
medicine.disease
biology.organism_classification
Invertebrates
030104 developmental biology
Antigens, Helminth
biology.protein
Immunologic Techniques
Enzymology
Serine Proteases
Developmental Biology
Subjects
Details
- Language :
- English
- ISSN :
- 19352735 and 19352727
- Volume :
- 12
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- PLoS Neglected Tropical Diseases
- Accession number :
- edsair.doi.dedup.....4de1c757f70e4eb0b170c8c25e4cf8f2