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M Parkhouse

Publications and source records attributed to M Parkhouse.

5 recordsLinked to original sources
PubMed2000

Cysticercosis: towards the design of a diagnostic kit based on synthetic peptides.

Cysticercosis caused by Taenia solium is a very common disease in developing countries that seriously affects human health. Diagnosis can only be confirmed with the aid of computerized tomography or nuclear magnetic resonance (NMR) creating obvious difficulties for epidemiological studies. Reliable immunoassays employing cerebrospinal fluid (CSF) have been developed, based on the use of cysticercal antigens. However, the reliance on parasite material is restrictive. Herein, we report the advances in the design of a diagnostic kit based on immunodominant synthetic peptides, targeting four candidate epitopes KETc1, KETc12, 410 and 413 which were identified from three different clones (KETc1, 12 and 4) selected from a cDNA library of Taenia crassiceps. CSF antibodies against T. solium cysticercal antigens (TCA) as well as the four peptides were determined by enzyme-linked immunoabsorbent assays (ELISA) using two panels of CSF from patients with confirmed neurocysticercosis and other neurological diseases. In the first CSF panel which included patients with high level of antibodies against TCA, KETc12 exhibited almost the same sensitivity (87.5%) as TCA (93.7%) and 100% specificity. In the second panel of 110 CSF collected at random, two peptides (KETc1 and KETc12) exhibited sensitivities of 40 and 36% respectively, and were 100% specific.

Animals↗
PubMed1998

A new membrane-bound OprI lipoprotein expression vector. High production of heterologous fusion proteins in gram (-) bacteria and the implications for oral vaccination.

We have previously described the development of cloning vectors for the production of OprI-based outer membrane fusion proteins in E. coli (Cornelis et al., 1996) and now describe the construction of a new vector, containing a lacI(q) gene, resulting in tight repression of the promotor and allowing its use in other Gram (-) bacteria. The new pVUB3 expression vector encodes a truncated but active LacI(q)(341) repressor which binds to the single operator in the vector. A high repression of the trc promotor was observed, resulting in a very low basal leakage of expression and very high production levels of OprI or derivatives after IPTG induction in E. coli. Bacterial viability was not affected under uninduced conditions, but the number of viable cell counts decreased after production of large amounts of the outer membrane-bound OprI lipoprotein and its derivatives, both in E. coli and Salmonella typhimurium. This highly repressible system allows us to extend the use of OprI vectors in other Gram (-) bacteria, resulting in the production of outer membrane-bound lipid-modified molecules, opening the possibility for its application in the design of potential live Salmonella-based subunit vaccines.

Amino Acid Sequence↗
PubMed1998

Diagnosis of porcine cysticercosis: a comparative study of serological tests for detection of circulating antibody and viable parasites.

Epidemiological studies of porcine cysticercosis require identification of pigs harbouring viable Taenia solium cysticerci and estimates of the degree of exposure to the parasite in the pig population destined for human consumption. Identification of infected pigs with viable larvae is achieved through detection of their secretory products. However, detectable levels of circulating antibody may also be present in the absence of viable larvae. In this study, both types of tests have been evaluated in groups of pigs experimentally infected with T. solium. Detection of viable cysticerci was achieved using a monoclonal antibody-based (HP10) antigen capture assay. HP10 epitope-bearing antigens have now been demonstrated in T. solium and T. crassiceps cyst fluid and excretion/secretions. Serum antibodies were measured in ELISA assays using two parasite preparations as antigens; T. solium cyst fluid and T. crassiceps cyst fluid antigens bearing the HP10 epitope. Low-background values were obtained with sera from non-infected animals in all the assays used. In heavily infected pigs, both antigens and antibodies were detected at least 29 days and up to 200 days post-infection (pi), while in lightly infected pigs antigen and antibodies were first observed between 61-97 days pi. Thus, the levels of the serum antigen and antibody varied with the intensity of the infection.

Animals↗
PubMed1978

[Identification of Entamoeba histolytica membrane antigens with antibodies of amebiasis patients].

The specificity of sera from patients with amoebiasis was assayed on trophozoites of E. histolytica HK9, HM15 and HM2 cultured axenically and monoxenically. Glutaraldehyde fixed cell were incubated with samples of serum, the excess of Igs was washed out and the presence of unbound material of specifically bound antibodies was measured with radiolabeled protein A from Staphylococcus aureus. A positive reaction with HK9 was observed in 18 per cent of the 66 sera tested. An immune crossreaction resulted with HM2 and HM15, while no reaction was observed with the nonpathogenic strains E. invadens, E. moshkovskii and E. laredo.

Amebiasis↗