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M D Curran

Publications and source records attributed to M D Curran.

52 records · Page 3Linked to original sources
PubMed1996

Long-range PCR amplification as an alternative strategy for characterizing novel HLA-B alleles.

We have developed a simple, rapid and reliable method for specifically amplifying and cloning full-length HLA-B genes from genomic DNA. Using this methodology we characterized three alleles of interest at the molecular level. Two of the alleles appeared in our routine class I PCR-SSOP typing system, a variant of B*5801 found in the Daudi cell line and RCE 56 and a variant of B*4101 found in a number of volunteer donors on our Bone Marrow Donor Registry. The third, a variant B35 allele found in RCE 80, was first identified as unusual by serology. Our sequencing analysis of exon 2 and exon 3 identified two of these alleles as the recently reported novel HLA-B*5802 and HLA-B*4102 alleles, while the third represents a new B35 allele officially designated B*3513.

Alleles↗
PubMed1995

Genetic diversity in the non-structural gene of parvovirus B19 detected by single-stranded conformational polymorphism assay (SSCP) and partial nucleotide sequencing.

A homologous region in the parvovirus B19 non-structural gene (B19 nt 1399-1682) was examined in 50 samples from patients with a wide variety of B19-related disease from various countries by PCR amplification, single-stranded conformational polymorphism (SSCP) assay and nucleotide sequencing. Five SSCP types were confirmed by nucleotide sequence analysis. Of a total of 6 mutations, all were silent. Types 3 and 4 accounted for 92% of strains. There was no correlation between genome type and either clinical illness or patient age. However, there was a correlation between SSCP type and country of origin. Type 3 strains predominated in Japan (18/26) and the UK (6/8), whereas type 4 predominated in the USA (9/12). Notably, type 3 strains also predominated among females (14/18), whereas there were approximately equal numbers of strain types 3 (7/17) and 4 (8/17) among males; an observation which remains unexplained. Within the Japanese group, although type 3 strains predominated overall, strains isolated from 1981 to 1987 consisted of types 1 (2/15), 2 (1/15), 3 (8/15), and 4 (4/15), whereas strains isolated from 1990 to 1994 consisted almost entirely of type 3 (10/11).

Adolescent↗
PubMed1995

Investigation of a nosocomial outbreak of Pseudomonas aeruginosa pneumonia in an intensive care unit by random amplification of polymorphic DNA assay.

From July to September 1993 in the intensive care unit of the Royal Victoria Hospital there were 10 cases of pneumonia associated with sputum culture of Pseudomonas aeruginosa. The isolates had an identical biotype and pyocine typing profile. The same strain of P. aeruginosa was recovered from the sink plug-hole in two rooms, and the tap handles and ventilator tubing in a third room. All strains were retrospectively typed by the random amplification of polymorphic DNA (RAPD) method using a 26-mer oligonucleotide primer, and were identical in profile. Recommendations to medical and nursing staff included secretion isolation precautions, terminal disinfection after patient discharge, use of disposable vinyl gloves by hospital staff for all body substance contacts, thorough handwashing with 4% chlorhexidine gluconate before and after dealing with all patient contacts, and prompt, appropriate antibiotic treatment for P. aeruginosa pneumonia. RAPD is a simple and effective method to determine the relatedness of P. aeruginosa isolates, and typing results are available within a single working day; thus dramatically increasing its clinical relevance over existing molecular methods.

Bacterial Typing Techniques↗
PubMed1995

A study of the role of parvovirus B19 in rheumatoid arthritis.

Serum and synovial tissue from 26 patients with rheumatoid arthritis (RA) (according to the diagnostic criteria of the American Rheumatism Association) and 26 patients with osteoarthritis (OA) were examined. Among the RA group, the female to male ratio was 4.2:1, and the age range was 44-82 yr with a mean of 64.0 yr; joints from which synovium was sampled were hip (n = 12), knee (n = 9), ankle (n = 3) and shoulder (n = 2). The duration of rheumatoid disease ranged from 6 to 24 yr with a mean of 13.9 yr. Among the OA group, the female to male ratio was 2.25:1, and the age range was 51-88 yr with a mean of 68.2 yr; joints from which synovium was sampled were hip (n = 18) and knee (n = 8). Twenty-one patients from the RA group and 20 patients from the OA group had evidence of previous parvovirus B19 infection (serum anti-B19 IgG), and all patients from both groups were serum anti-B19 IgM negative. Synovial sections from all 52 patients were stained with mouse monoclonal antibodies, 3H8 (to B19 capsid proteins) and alpha-P (to blood group P antigen). All tissue sections examined were found to be negative for both B19 capsid proteins and blood group P antigen. Using a nested polymerase chain reaction (PCR) assay, all patients were negative for serum B19 DNA. However, B19 DNA was demonstrated in the synovium of 10 of 26 RA patients and 9 of 26 OA patients; uncorrected chi 2 value = 0.08; degrees of freedom = 1; P = 0.77. All 19 patients testing positive for synovial B19 DNA had evidence of prior exposure to B19 infection (serum anti-B19 IgG). In conclusion, although there is published evidence of chronic rheumatoid-like arthropathy following acute parvovirus B19 infection, our findings do not support the involvement of B19 in the aetiopathogenesis of RA.

Adult↗
PubMed1995

Parvovirus B19 infection--persistence and genetic variation.

53 patients with acute B19 infection were studied; symptoms at acute infection were rash and arthralgia (n = 26), rash (n = 7), arthralgia (n = 16), aplastic crisis (n = 3), and intrauterine fetal death (n = 1). These patients were followed for 26-85 months (mean 57 months) and re-assessed for persistent symptoms, anti-B19 antibodies, and B19 DNA. At follow-up, 7 individuals were positive for serum B19 DNA, compared with none of the controls (2-tailed p value = 0.016). All 7 of those persistently infected were women, 3 of whom had symptoms; 1 had a chronic haemolytic anaemia (initial presentation was aplastic crisis); 1 had persistent arthralgia in both knees (initial presentation was bilateral knee arthralgia); and 1 had arthralgia in one knee and chronic fatigue syndrome (initial presentation was bilateral arthralgia in knees and shoulders). For the 7 persistently infected patients, serum from the time of diagnosis of acute B19 infection was available for 4, all of which contained B19 DNA. With single-stranded conformational polymorphism (SSCP) assay of these 11 PCR products, identical SSCP types were demonstrated in 5 of 7 follow-up isolates. In 2 of the 4 cases for which both acute and follow-up PCR product was available, the SSCP type of the follow-up product was different from that of the acute product. Two B19 virus types were demonstrated in one patient (with persistent arthralgia and chronic fatigue syndrome) at follow-up assessment.

Adolescent↗
PubMed1992

Phocine distemper virus, the agent responsible for the 1988 mass mortality of seals.

The biochemical characterisation of phocine distemper virus (PDV) has shown that PDV is related to but clearly distinct from canine distemper virus (CDV) and relative to its relationship with CDV is only remotely related to the other morbilliviruses, namely measles virus (MV) or rinderpest virus (RPV) and peste-des-petits-ruminants virus (PPRV). Comparative studies with monoclonal antibodies indicate that the virus is serologically closely related to CDV with many conserved epitopes, particularly on the internal proteins of the virus, while the external attachment (H) protein shows the greatest level of variability among the distemper virus isolates. The analysis of the viral proteins by electrophoresis indicates molecular weight differences between CDV and PDV in the fusion (F), phosphoprotein (P), H, nucleocapsid (N) and matrix (M) proteins. The RNA profiles of CDV and PDV are indistinguishable and different from those for RPV and MV. Nucleotide sequence analysis of cDNA clones of the virus show approximately 70% homology between CDV and PDV and approximately 48% with MV. These data prove that PDV is a different virus from CDV and co-circulates with it probably primarily in sea mammals.

Animals↗
PubMed1992

The fusion protein gene of phocine distemper virus: nucleotide and deduced amino acid sequences and a comparison of morbillivirus fusion proteins.

The nucleotide sequence of the gene encoding the fusion protein of phocine distemper virus has been determined. The mRNA is 2206 nucleotides in length and contains one major open reading frame (ORF) of 1893 nucleotides encoding a potential protein of 631 amino acid residues. However, analogy with canine distemper virus (CDV) suggests that translation of the F protein starts at the sixth AUG codon in the mRNA sequence which is located at position 461, resulting in an F0 protein of exactly the same size (537 aa) as that of CDV. The overall homology at nucleotide level between the CDV and PDV F genes is 66%. The homology between the two F proteins of these respective viruses is 83%.

Amino Acid Sequence↗
PubMed1992

Molecular characterization of phocine distemper virus: gene order and sequence of the gene encoding the attachment (H) protein.

Generation of a set of cDNA clones covering the N, P/V/C, M, F, H and part of the L gene of phocine distemper virus (PDV) has been described. The gene order of PDV determined from a physical as well as a transcriptional map, was identical to that of the other morbilliviruses so far studied. The H gene sequence (1951 nucleotides) contains one large open reading frame which encodes a protein of 607 amino acids, identical in length to that of the H protein of the Convac strain of canine distemper virus (CDV). Nucleotide and protein sequence comparisons between PDV and the other morbilliviruses provide further evidence in favour of PDV's inclusion in the morbillivirus genus as a distinct species closely related to CDV and more distantly to measles virus and rinderpest virus.

Amino Acid Sequence↗
PubMed1992

The genes encoding the phospho- and matrix proteins of phocine distemper virus.

The nucleotide sequences of the phosphoprotein (P)/V/C and matrix (M) protein genes of phocine distemper virus (PDV) have been determined and the deduced amino acid sequences of the proteins derived from these genes compared with those of the other morbilliviruses. The 1655 nucleotides of the P gene encode a phosphoprotein of 507 amino acid residues (from nucleotide numbers 60 to 1583) which is 75% identical to that of canine distemper virus (CDV). The C proteins of the two viruses are 73% identical. The C protein has the same length, 174 amino acid residues, as C of CDV. The nucleotide sequences of the P genes are 78% identical. The editing site in the P gene is present as a stretch of 18 nucleotides, conserved in all other morbilliviruses. A V-like protein can be accessed by insertion of one G residue at this site. The P and M genes are separated from adjacent genes and each other by the CTT trinucleotide sequence which is totally conserved in morbilliviruses. The M gene is 1447 nucleotides long and encodes a typical paramyxovirus M protein of 335 amino acids (identical in length to those of CDV, measles virus and rinderpest virus). The gene is 78% identical to CDV in the coding region (nucleotides 32 to 1039) but only 34% identical to CDV in the 407 nucleotides of the 3' untranslated sequence of the M mRNA. The M protein of PDV is 91% identical to the M protein of CDV. The data demonstrate interesting variations in the level of conservation of various genome areas and prove the distinct nature of PDV.

Amino Acid Sequence↗
PubMed1991

The nucleotide sequence of the gene encoding the attachment protein H of canine distemper virus.

The sequence of the H gene and flanking sequences in the F and L genes of canine distemper virus (CDV) have been determined. The H gene of CDV (1946 nucleotides) contains one large open reading frame starting at position 21 and terminating at position 1835, encoding a protein of 604 amino acid residues. This protein contains three potential glycosylation sites in the extracellular domain and, like all other paramyxoviruses, a N-terminal membrane-spanning hydrophobic anchor domain. The deduced H protein sequence shows an identity of 36% with rinderpest virus (RPV) and measles virus (MV). The identities at the nucleotide level are higher (RPV 52% and MV 53%). The amino acid sequence shows conservation of all the structural determinants with the H proteins of MV and RPV. The data also show that CDV is evolutionarily equidistant to RPV and MV with respect to the H gene.

Amino Acid Sequence↗
PubMed1988

Nucleotide sequence of the gene encoding the matrix protein of a recent measles virus isolate.

The sequence of the M gene of the Hu2 strain of measles virus has been determined. In the coding region of the gene, six nucleotide replacements had occurred with respect to the sequence of the M gene of the Edmonston strain. Two of these were silent and only four led to amino acid replacements in the protein sequence even though in SDS-PAGE the mobility of the protein would indicate an Mr increase of 2000. No changes were observed in the 5' untranslated regions of the gene. Two changes were observed in the 3' untranslated region of the gene. The sequences are compared to those recently published for subacute sclerosing panencephalitis viruses.

Amino Acid Sequence↗
PubMed1986

Characterization of clones for the sixth (L) gene and a transcriptional map for morbilliviruses.

cDNA clones of the largest RNA transcript of the canine distemper and measles morbilliviruses were characterized. This presumably codes for the L protein of these viruses. mRNA 4 was identified as coding for the haemagglutinin protein of measles virus. From an analysis of readthrough transcripts representing tandem copies of two or three genes we established a transcriptional map and the gene order on the negative strand genome of the morbilliviruses to be 3'-N-P + C-M-F-H-L-5'. The data exclude the presence of small intervening genes between the six major genes of morbilliviruses and indicate the gene order to be similar to that of Sendai virus and different from that of simian virus 5.

Cloning, Molecular↗
PubMed1985

Characterization of phages derived from strains of Rhodococcus australis and R. equii.

Rhodococcus australis strain CSIR-A201 and R. equii strain CSIR-A655 spontaneously liberated phages A and B, respectively. The phages have different host ranges, but both infect R. rubropertinctus ATCC 14352. phage infection of strains resulted in stable lysogeny. The phages have similar morphologies and belong to the family Styloviridae. The DNA restriction enzyme patterns of the phages differ and do not correspond to those of actinophage phi EC. These temperate phages did not transduce prototrophic or antibiotic resistance markers to appropriate hosts.

Actinomycetales↗