Strategy for identification & characterization of Bartonella henselae with conventional & molecular methods.

BACKGROUND & OBJECTIVES: Bartonella henselae is a fastidious gram-negative bacterium usually causing self limiting infections in immunocompetent individuals but often causes potentially life threatening infection, such as bacillary angiomatosis in immunocompromised patients. Both diagnosis of infections and research into molecular mechanisms of pathogenesis have been hindered by lack of appropriate and reliable diagnostic techniques. We undertook this study to standardize methods to characterize B. henselae in clinical samples to diagnose Bartonella infection correctly. METHODS: B. henselae ATCC 49882 strain was procured from American type culture collection, USA. This strain was revived and maintained in the laboratory, and identification and characterization of this strain was done by conventional and molecular techniques, which included culture on various media, staining by different methods including electron microscopy, biochemical analysis by conventional methods and API, polymerase chain reaction (PCR) for amplification of citrate synthase gene followed by restriction fragment length polymorphism (RFLP). RESULTS: This organism was biochemically inert due to slow growth and generated unique identification code with API. The amplification of the citrate-synthase gene with primers yielded a 381 bp product followed by specific RFLP profile for B. henselae. INTERPRETATION & CONCLUSIONS: Bartonella is fastidious and fragile organism and should be handled carefully. Extra effort and careful observation are required to isolate and characterize this organism.

Institutional outbreak of rubella in a healthcare center in Chandigarh, North India.

Rubella is traditionally considered a childhood disease but it has the potential to cause outbreaks in closed communities when a susceptible population accumulates. The present study reports an outbreak of rubella among healthcare workers in the pediatric center of a tertiary care North Indian hospital. The cases of rubella were identified by clinical features and confirmed by the detection of anti-rubella IgM antibodies in blood by enzyme linked immunosorbent assay. A total of 23 cases of rubella occurred over a period of one and a half month, out of which 9 (39%) were males. All the patients were in the age group of 21-35 years. None of the patients gave a history of rubella vaccination. This outbreak of rubella occurred due to the accumulation of a susceptible population in a closed hospital environment. There is need for the introduction of rubella vaccination in healthcare workers to prevent outbreaks at work place.

Concurrent infections by all four dengue virus serotypes during an outbreak of dengue in 2006 in Delhi, India.

BACKGROUND: Co-circulation of multiple dengue virus serotypes has been reported from many parts of the world including India, however concurrent infection with more than one serotype of dengue viruses in the same individual is rarely documented. An outbreak of dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) occurred in and around Delhi in 2006. This is the first report from India with high percentage of concurrent infections with different dengue virus serotypes circulating during one outbreak. RESULTS: Acute phase sera from patients were tested for the presence of dengue virus RNA by RT-PCR assay. Of the 69 samples tested for dengue virus RNA, 48 (69.5%) were found to be positive. All the four dengue virus serotypes were found to be co-circulating in this outbreak with DENV-3 being the predominant serotype. In addition in 9 of 48 (19%) dengue virus positive samples, concurrent infection with more than one dengue virus serotype were identified. CONCLUSION: This is the first report in which concurrent infections with different dengue virus serotypes is being reported during an outbreak from India. Delhi is now truly hyperendemic for dengue.