• PDF / 171,144 Bytes
• 1 Pages / 595.245 x 841.846 pts (A4) Page_size
• 19 Downloads / 219 Views

DOWNLOAD

REPORT

---

1 S

Disseminated BCG infection: case report An approximately 14-month-old girl developed disseminated BCG (Bacillus Calmette-Gu´erin) infection after receiving BCG vaccination. The girl, at 12 months of age, underwent bone marrow transplantation for severe combined immunodeficiency (SCID). At that time, busulfan and fludarabine were used as conditioning regimen. Ciclosporin [cyclosporine] and mycophenolate mofetil were used as prophylaxis for graft-versus-host disease. After 2 months, she developed skin ulceration, fever, hepatomegaly, vomiting and failure to thrive. On initial investigation, no lymphadenopathy and bone involvement were identified. Later, abdominal ultrasound demonstrated hepatomegaly, mesenteric lymphadenopathy, and ascites and infiltration around intestinal loops. Also, pathologic examination of a liver biopsy revealed granulomatosis, including multinuclear giant cells with a lymphocytic crown. Thereafter, RTPCR of skin and mesenteric lymph node biopsies were found to be positive for rifampicin-susceptible Mycobacterium tuberculosis complex. At that time, she was diagnosed with systemic tuberculosis. Therefore, the girl started receiving treatment with isoniazid, rifampicin, amikacin ethambutol and ciprofloxacin combinedly. Later, at a 10 months follow-up, she was afebrile, clinically well with 1.4kg weight gain and normal biology. At that time, the vaccination notebook revealed that, at 3 days of age, she had received intradermal Russian BCG vaccine [Moscow–368] 0.05mg. At that time, the previously stored samples of biopsy specimens were triturated and inoculated on human embryonic lung fibroblasts and on endothelial cells using the shell-vial assay. After 11 days of co-culture, microscopy with Ziehl-Neelsen staining demonstrated mycobacteria in both the cell lines. Thereafter, the total DNA of mycobacteria bovis BCG was extracted from the positive cell cultures and sequenced. Paired-end sequencing and automated cluster generation of 5 runs, with dual-indexed 2 x1 50-base pair reads, were performed for 17.5 hours. An online tool, TGS-TB (total genotyping solution for mycobacteria tuberculosis) identified the Mycobacterium, which was confirmed subsequently. Genomic reads were then mapped to one of the Mycobacterium BCG Pasteur 1173P2 complete genomes. An extracted consensus sequence yielded 451 contigs with 4,333,926 case pair and 65.5% GC content. An annotated genome, using Prokka version 1.12, yielded 4272 predicted genes, including 4216 protein-coding genes, 2 repeat regions and 56 RNA genes, including 52 tRNA, 3 rRNA and 1 tmRNA. In silico whole-genome analysis noted a natural resistance to pyrazinamide (H57D pncA gene) and susceptibility to rifampicin, isoniazid, ethambutol, ciprofloxacin and amikacin. Based on these findings, at the 19 months of age, she was diagnosed with disseminated BCG infection. Fellag M, et al. Shell-vial Assay in Diagnosis of Disseminated BCG Infection in an Immunodeficient Child. Pediatric Infectious Disease Journal 39: 258-259, No. 3, Mar 803520093 2020. Available