Screening for calreticulin mutations in a cohort of patients suspected of having a myeloproliferative neoplasm.

BACKGROUND: The discovery of calreticulin (CALR) has shown it to be the second most frequent mutation after the Janus Kinase 2 (JAK2) mutation in myeloproliferative neoplasms (MPNs). Its structure indicates various functions, of which two are to ensure calcium homeostasis and proper folding of other target proteins. Over 36 types of CALR mutations have been identified, all causing a recurrent frameshift in the C-terminal domain affecting CALR's localisation and calcium-binding function. OBJECTIVE: To screen a cohort of 89 patients suspected of having an MPN for the CALR mutations. METHODS: Capillary and gel electrophoresis were used in conjunction as confirmatory tests to screen the cohort of patients. RESULTS: Of three samples containing a type 1 CALR mutation, two were heterozygous and one homozygous for a 52-base pair deletion in CALR. CONCLUSIONS: Most studies report CALR mutations to be present only in patients with primary myelofibrosis or essential thrombocythaemia, with mutual exclusivity to JAK2 mutations. The findings of this study indicate that JAK2 and CALR mutations are no longer considered mutually exclusive. Similarly, patients with a polycythaemia vera phenotype could also carry a CALR mutation.

Adult-to-child transmission of tuberculosis: household or community contact?

SETTING: If a child develops tuberculosis, it is assumed that the source was an adult infectious case, usually living in the same house. Restriction fragment length polymorphism (RFLP) was used in this study to establish transmission from source cases to children. DESIGN: Adult and child tuberculosis cases were prospectively identified from 1993 to 1998 and cultures of Mycobacterium tuberculosis collected. Interviews and RFLP analysis of M. tuberculosis strains were performed to establish epidemiological links and to confirm household transmission. RESULTS: Tuberculosis was confirmed by culture in 1139 (91%) of 1291 adults and 65 (16%) of 417 children. Due to problems in recovering specimens or extracting DNA, RFLP analysis was done in 832 adults and 35 children: 19 (54%) children had household members identified with tuberculosis, 12 with the same strain as the child. Twenty-nine (83%) strains from children formed part of community clusters, but definite contact with source cases was established in only 15. CONCLUSION: The presence of an adult with infectious tuberculosis in the same house as a child with tuberculosis does not necessarily imply adult-to-child transmission. Young children may be infected in the community or in the household. These findings have implications for contact tracing and treatment strategies in high incidence areas.

Tuberculosis transmission patterns in a high-incidence area: a spatial analysis.

SETTING: In the Cape Town suburbs of Ravensmead and Uitsig, tuberculosis has reached epidemic levels, with notifications of 1340/100,000 in 1996. These suburbs are characterised by overcrowding, high unemployment and poverty. It is traditionally believed that tuberculosis transmission takes place mainly in households after close contact with an infectious person. Studies have recently linked tuberculosis transmission to locations outside the household, and have associated these places with a particular high-risk lifestyle. Anthropological studies in some suburbs of Cape Town, in which a very high number of local drinking places (shebeens) were identified (17 per km2), have suggested that social drinking is part of such a lifestyle. OBJECTIVE: To investigate various risk factors and places of transmission of tuberculosis using a geographical information system (GIS). RESULTS AND CONCLUSION: The 1128 bacteriologically-proven cases of tuberculosis studied over the period 1993-1998 were investigated using spatial epidemiological techniques of exploratory disease mapping. Point pattern analysis and spatial statistics indicated clustering of cases in the areas of high incidence. Significant associations of tuberculosis notifications were found with unemployment, overcrowding and number of shebeens per enumerator sub-district. High tuberculosis notifications with unemployment and its associated poverty emerged as the strongest association.

Historic and recent events contribute to the disease dynamics of Beijing-like Mycobacterium tuberculosis isolates in a high incidence region.

SETTING: A retrospective study in an urban setting with a high tuberculosis incidence. OBJECTIVE: To study the molecular epidemiology and disease dynamics of a prevalent Mycobacterium tuberculosis strain family, F29, a subset of the Beijing strains. DESIGN: Multi-locus DNA fingerprinting and genomic analysis was used to characterise F29 isolates. Demographic and molecular epidemiologic data were correlated with those of other isolates from the setting. The geographic locations of five F29 strain clusters were plotted using a geographic information system (GIS) and an index for geographic distribution was calculated. Their cumulative temporal occurrence was also plotted. RESULTS: The genomic similarity of F29 to the Beijing-strains was confirmed. A high degree of clustering predicted high disease transmission. Spatial distribution was mostly widespread except for one cluster. Smaller foci of transmission were observed. The temporal spread showed ongoing transmission. CONCLUSION: F29 belongs to the Beijing strain group. The prevalence and high degree of strain clustering, with limited geographic clustering, indicates that F29 strains are endemic in the study community. However, recent epidemiologic events also contributed to the disease spread. The combination of molecular epidemiologic, spatial and temporal data has enhanced our understanding of the disease dynamics of Beijing strains in our study community.

Evolution of the IS6110-based restriction fragment length polymorphism pattern during the transmission of Mycobacterium tuberculosis.

Interpretation of the molecular epidemiological data of Mycobacterium tuberculosis is dependent on the validity of the assumptions that have been made. It is assumed that the IS6110 banding pattern is sufficiently stable to define epidemiological events representing ongoing transmission. However, molecular epidemiological data also support the observation that the IS6110 banding pattern may change over time. Factors affecting this rate may include the nature and duration of disease in a host and the opportunity to experience different host environments during the transmission cycle. To estimate the rate of IS6110 change occurring during the process of transmission, M. tuberculosis isolates from epidemiologically linked patients were genotypically characterized by restriction fragment length polymorphism (RFLP) analysis. The identification of IS6110 banding pattern changes during ongoing transmission suggested that a rate could be estimated. IS6110 change was significantly associated with strains with >5 IS6110 elements (P = 0.013) and was not observed in low-copy-number isolates. The minimum rate of appearance of variant strains was calculated to be 0.14 variant cases per source-case per year. This data suggest that clustering of isolates based on identical RFLP patterns is expected to underestimate transmission in patients infected with high-copy-number isolates. A model based on the rate of appearance of both variant and invariant strains demonstrates that the genotypically defined population structure may change by 18.6% during the study period of approximately 6.5 years. The implications for the use of RFLP data for epidemiologic study are discussed.