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C McCaughey

Publications and source records attributed to C McCaughey.

16 recordsLinked to original sources
PubMed2001

Nested multiplex polymerase chain reaction for the diagnosis of cutaneous herpes simplex and herpes zoster infections and a comparison with electronmicroscopy.

Herpes simplex virus (HSV) and varicella zoster virus (VZV) are common causes of cutaneous and mucocutaneous vesicular eruptions. Laboratory diagnostic techniques include Tzanck smears, electronmicroscopy, antigen detection and viral culture. This paper describes a nested multiplex polymerase chain reaction with respective sensitivities of 0.0001, 0.01 and 0.1 TCID50 for VZV, HSV-1 and HSV-2. The assay was used in (a) a salvage capacity for slides already processed for electronmicroscopy, and (b) as a front-line assay for prospectively processed specimens. Sixty-two glass slides with vesicle lymph/scrapings from 58 patients with suspected cutaneous herpetic lesions were examined. The clinical presentations were described as atypical/not specified (24), VZV (20) or HSV (18), and involved eruptions from diverse anatomical sites, including the genitalia. Of the 62 specimens, 6 and 38 were positive by electronmicroscopy and multiplex PCR respectively, giving a comparative sensitivity of 16% for electronmicroscopy. Nested multiplex PCR identified 15 VZV and 20 HSV-1 infections. Where the clinical details indicated either HSV or VZV (38/62), nested multiplex PCR was statistically likely to be reactive (26/38 vs. 9/24) (chi2 P = 0.000004) whereas electronmicroscopy was not (4/38 vs. 2/24) (chi2 P= 0.77). Where the clinical details indicated VZV (20/62) or HSV (18/62), nested multiplex PCR was statistically more likely to confirm VZV (10/20 vs. 5/42) (chi2 P= 0.001) or HSV (9/18 vs. 11/44) (chi2 P = 0.05) respectively. Two suspected HSV and 6 suspected VZV infections were shown to be VZV and HSV respectively by nested multiplex PCR.

Adolescent↗
PubMed1999

Low pH-induced cytopathic effect--a survey of seven hantavirus strains.

Hantaviruses do not produce cytopathic effects (CPE) in cell culture. However, a syncytial CPE can be induced in 7-day cultures of hantavirus growing in Vero E6 cells by reduction of the pH to approximately 6.2 using a HEPES based buffer. The appearance of this acid induced CPE was examined for seven different hantavirus strains. The differences noted were striking and reflected the taxonomic differences between hantaviruses. At 10-100 TCID50% the size of syncytial foci was very large for Seoul type viruses and smallest for Puumala viruses. The size of syncytia for Hantaan (HTN) virus was intermediate between Puumala (PUU) and Seoul (SEO) type viruses.

Animals↗
PubMed1999

A comparison of virus isolation, indirect immunofluorescence and nested multiplex polymerase chain reaction for the diagnosis of primary and recurrent herpes simplex type 1 and type 2 infections.

134 swabs in viral transport medium were received from 126 patients with suspected clinical HSV-1 and HSV-2 infections. They were tested by (i) nested multiplex polymerase chain reaction NMPCR (strongly positive specimens had visible bands on both rounds of PCR) without prior extraction, (ii) culture in primary rhesus monkey kidney, E6-Vero, RD and HEp-2 cells and (iii) antigen detection by immunofluorescence (IF). Antigen detection employed four novel pools (A-D) of monoclonal antibodies (Mab): A was HSV-1 specific, B was HSV-2 specific while C and D were generic. In comparison to NMPCR the sensitivity and specificity of (i) culture was 59% (22/37) and 100% (134/134), (ii) IF by Pool A was 59% (16/27) and 100% (117/117), (iii) IF by Pool B was 40% (4/10) and 100% (130/130) and (iv) IF by Pools C and D were 60% (18/30) and 100% (96/96). Specimens positive by culture were more likely to be strongly positive by NMPCR (chi2 P = 0.004). Typing by each method concurred on all occasions. NMPCR was cost effective, easier to perform and was the most sensitive method for HSV detection. It should become the method of choice for HSV diagnosis.

Base Sequence↗
PubMed1998

Effect of faecal consistency on virological diagnosis.

A study was set up to investigate the effect of consistency of routine faecal specimens on the diagnostic yield by electron microscopy (EM) and virus isolation. A total of 3078 specimens were characterized as solid, semisolid, or liquid. Of 2568 specimens processed by EM a virus was demonstrated in 8.6% of liquid, 19.9% of semisolid and 25.2% of solid specimens (Chi-squared for linear trend, P value <0.0001). This observation was valid for both adenovirus (2.4%, 5.0% and 6.6%) and rotavirus (5.2%, 13.6% and 16.6%). Virus isolation was positive in 3.6% of liquid, 17.4% of semisolid and 18.1% of solid specimens. (Chi-squared for linear trend, P value <0.0001). We suggest that solid faecal specimens at the end of an episode of diarrhoea will have a higher diagnostic yield than liquid specimens at the peak of symptoms. Our findings repudiate the commonly held dogma that viruses of gastroenteritis are more likely to be found in liquid than in solid faecal specimens. This finding has important implications for those establishing diagnostic algorithms for the investigation of viral gastroenteritis.

Adenoviridae↗
PubMed1998

Nucleotide sequence and phylogenetic analysis of the medium (M) genomic RNA segments of three hantaviruses isolated in China.

The medium (M) genome segment of hantaviruses (family Bunyaviridae) encodes the two virion glycoproteins. G1 and G2, as a precursor protein in the complementary sense RNA. We determined the nucleotide sequences of the M genome segments of three Chinese hantavirus isolates, a Hantaan-type (HTN) virus designated A9 and two Seoul-type (SEO) viruses designated L99 and HB55, and compared them to those of other HTN or SEO viruses isolated in Eastern Asia. The M segment of A9 is 3616 nucleotides in length and shows 99.5% identity at the nucleotide level and 99.1% identity at the amino acid level to that of the Chinese HTN isolate HV114. The M segments of L99 and HB55 are 3652 nucleotides in length, one nucleotide longer than the M segments of other sequenced SEO isolates such as SEO 80-39, SR-11, and Biken-1. The Chinese SEO isolates showed 95% nucleotide sequence identity and 99% amino acid sequence identity to SEO 80-39. We also sequenced a 736 nucleotides region of the M genome segment of another Chinese SEO isolate, R22, which revealed errors in the published data. Phylogenetic analysis of the available sequences indicated that both the Chinese HTN- and SEO-type viruses form lineages distinct from those of the isolates from other parts of Eastern Asia.

Amino Acid Sequence↗
PubMed1998

Recrudescent herpes simplex infection mimicking primary herpetic gingivostomatitis.

We report six cases of recrudescent intraoral herpes simplex infection clinically indistinguishable from primary herpetic gingivostomatitis. All infections occurred in healthy children or young adults. Serological analysis demonstrated herpes simplex virus (HSV)-specific IgG at initial presentation, indicating that the infection was not a primary infection. Convalescent sera exhibited HSV-specific IgM and a rising HSV-specific IgG titre. These findings demonstrate that the initial clinical diagnosis of primary herpetic gingivostomatitis was erroneous and that what was actually being observed was widespread recrudescent intraoral herpes simplex infection.

Adolescent↗
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↗