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Biomedical subjects

C McCaughey

Publications and source records attributed to C McCaughey.

26 records · Page 2Linked to original sources
PubMed1996

Isolation of viruses from clinical specimens in microtitre plates with cells inoculated in suspension.

Virus isolation is essential for the provision of a full diagnostic virology service. Present methods are time consuming, expensive and relatively inflexible for routine use. Our objective was to audit our existing virus isolation system and to develop a sensitive, flexible virus isolation system which could be adapted for use in a busy routine laboratory which is required to provide a service for a wide range of clinical situations. We carried out a pilot study which compared conventional roller tube monolayer cultures to a microplate system using cells inoculated in suspension and showed that the microplate method using extra cell lines could provide a more sensitive system for virus isolation. This system was adapted for routine use using six cell lines inoculated in suspension and the results are presented for 2610 specimens for virus isolation and 972 for Clostridium difficile toxin (CDT) detection. There were 516 viruses isolated and 229 specimens positive for CDT using this system. Polioviruses (92), echoviruses (35), coxsackieviruses (15) and untyped enteroviruses (13) were isolated in RMK, E6-vero and RD cells. Adenoviruses (137) were isolated in HEp2 and E6-vero cells. Herpes simplex virus (HSV) was isolated from 149 specimens in E6-vero, FCL and HFF9 cells. Myxoviruses (38) and paramyxoviruses were isolated in RMK cells. HEp2 was the only cell line necessary to isolate the 33 respiratory syncytial viruses (RSV). Cytomegaloviruses (CMV) (2) and varicella zoster (1) virus (VZV) were isolated only in the human fibroblast cell line HFF9. Rubella virus was isolated from a baby with congenital rubella in RMK, E6-vero and additionally in BGM cells. In conclusion, the use of cells inoculated in suspension in microtitre plates for virus isolation was sensitive and convenient. It allowed the use of six cell lines for routine virus isolation without using additional laboratory staff time. It improved turnaround times. It was also safer microbiologically than conventional isolation in tube monolayers. The precise identification of virus isolates was simplified.

Adenoviridae↗
PubMed1996

Evidence of hantavirus in wild rodents in Northern Ireland.

A survey of evidence of rodent hantavirus infection in County Down, Northern Ireland was carried out by using immunofluorescence to detect virus antigen and antibody. Antibodies to hantavirus (R22 strain of Seoul virus and Hantaan 76-118) were found in 11/51 (21.6%) brown rats (Rattus norvegicus), 1/31 (3.2%) field mice (Apodemus sylvaticus) and 17/59 (28.8%) house mice (Mus domesticus). Seven rodents had evidence of hantavirus antigen in lung tissues. Antibody positive animals were significantly more likely to be adults than juveniles (P = 0.04) but and there was no sex difference between antibody positive and negative animals. House mice were more likely to be antibody positive if captured inside farm outbuildings (P = 0.08). Attempts to culture virus from the rodent material were unsuccessful. This work demonstrates a substantial rodent reservoir for hantavirus in Northern Ireland.

Animals↗
PubMed1994

Serological evidence of Hantavirus disease in Northern Ireland.

Since, to our knowledge, no clinically documented cases of haemorrhagic fever with renal syndrome (HFRS) have been reported in Northern Ireland, a sero-epidemiological study was carried out to assess the degree of Hantavirus immunity in a group of 627 Northern Irish patients presenting with symptoms suggestive of HFRS and 100 healthy controls. IFA screening for IgG Hantavirus specific antibodies was carried out with a panel of up to 9 different Hantaviral antigens. IgM screening was performed using a commercially available mu-capture ELISA based upon two recombinant Hantaviral nucleocapsid antigens. A seropositivity of 2.1% (15/727) was recorded, with an almost exclusive reaction against a rat-derived R22VP30 strain of the Seoul serotype. Sole reliance upon non-rat-derived classic screening antigens Hantaan (HTN 76-118) and Puumala (CG 18-20) would have resulted in the detection of only 2/15 (13.3%) of cases in IgG IFA, and 8/14 (57.1%) of the cases in IgG ELISA. The findings indicate that for the first time in Europe, and more specifically in N. Ireland, non-laboratory outbreaks of HFRS may be caused by wild rats acting as a reservoir for a Seoul-like Hantavirus. Conventional Hantavirus serology using Hantaan and Puumala as screening antigens does not appear sufficient for the detection of such cases of HFRS. Hence, we propose the addition of a rat derived Hantaviral antigen to the antigen screening panel as a means of improving the specificity of the detection methods.

Animals↗
PubMed1992

A simple standardised protocol for the production of monoclonal antibodies against viral and bacterial antigens.

A simple standardised protocol for making monoclonal antibodies against a range of human bacteria and viruses is described. The protocol was designed to reduce the number of steps to a minimum. A one step footpad immunisation was followed by the fusion schedule 10-15 days later. A vital step in the technique was the use of the immunised mouse's spleen to provide a feeder layer post fusion. This simplified the protocol and more importantly greatly accelerated the growth of the hybridomas produced. Immunisation, fusion and clonal expansion of specific antibody secreting hybridomas was complete within 5 weeks. The percentage of hybridomas secreting specific antibody ranged from 6% to 28%, the majority of which were of the IgG isotypes. The method was economical in the use of tissue culture medium and simple to perform.

Animals↗