Identification of substrains of BCG vaccine using multiplex PCR

Bedwell, J.; Kairo, Satnam K.; Behr, Marcel A.; Bygraves, Jane A.

doi:10.1016/s0264-410x(00)00369-8

Cited by 69 publications

(69 citation statements)

References 15 publications

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“…Although in the majority of countries with high TB incidence, the vaccine is supplied by UNICEF/WHO and the Global Alliance for Vaccines and Immunization (GAVI); in Iran as one of those countries with high TB incidence, a local unknown BCG vaccine strain is being used (37). For vaccine production purposes, different passage conditions have been used in different laboratories between 1921 and 1961, which have led to the emergence of different BCG vaccine strains, with genetic variations, including deletions, duplications and point mutations (37,38). These genetic events can affect the immunogenicity of BCG and therefore ineffective immune response against BCG vaccine.…”

Section: Resultsmentioning

confidence: 99%

The History of Tuberculosis and Bacillus Calmette–Guérin Vaccine in Iran

Fallah

Abdolghafoorian

2014

Arch Pediatr Infect Dis

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Section: Resultsmentioning

confidence: 99%

The History of Tuberculosis and Bacillus Calmette–Guérin Vaccine in Iran

Fallah

Abdolghafoorian

2014

Arch Pediatr Infect Dis

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“…For the purpose of quick and accurate identification of the sample, we ran multiplex PCR on the sample by a simple modification of the method of Bedwell et al (1). In 1999, a DNA microarray clarified that 16 regions in BCG strains, named RD1 to RD16, were different from those in M. tuberculosis H37Rv and that some regions differed among BCG strains (2).…”

Section: Bovis)mentioning

confidence: 99%

“…Since PCR products for SenX3-RegX3 of M. bovis, except BCG strains, and those for RD16 of BCG Tokyo 172 were very close in size, we modified the original multiplex PCR method of Bedwell et al by simply excluding primers for RD16. Thus, our multiplex PCR contains primers for RD1, the complement sequence of RD1, RD2, RD8, RD14, and SenX3-RegX3, which were designed as previously described (1).…”

Section: Bovis)mentioning

confidence: 99%

“…The multiplex PCR amplification was performed as previously described (1). M. tuberculosis Aoyama B (obtained from the National Institute of Infectious Diseases, Tokyo, Japan) and M. bovis Ushi 10 (obtained from the Japanese Association of Veterinary Biologics) showed a 150-bp RD1 product, whereas both M. bovis BCG Tokyo 172 and the colony showed 200-bp bands as the complement sequence of RD1, indicating the deletion of RD1 (Fig.…”

Section: Bovis)mentioning

confidence: 99%

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Multiplex PCR-Identified Cutaneous Tuberculosis Evoked by Mycobacterium bovis BCG Vaccination in a Healthy Baby

et al. 2005

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This is the first identified case of Mycobacterium bovis bacillus Calmette-Guérin (BCG)-derived cutaneous tuberculosis that localizes at a place different from the vaccination site in hosts without immune deficiency. A healthy baby with a developing abscess is described. A multiplex PCR identified the abscess as originating from M. bovis BCG Tokyo 172. CASE REPORTA 6-month-old girl with no previous significant medical problems developed an abscess in her left humerus. She had been vaccinated with Mycobacterium bovis bacillus CalmetteGuérin (BCG) Tokyo 172 at the age of 4 months. At 5 months, a red and swelling nodule appeared in her left humerus at a place different from the vaccination site. At 6 months, she visited our hospital. A red, fluctuating, and swollen abscess about 2 cm from the region of the vaccination was found. The abscess measured 10 by 15 mm. We found no swelling of the lymph nodes. The patient was afebrile and did not have respiratory symptoms. There was no abnormal chest radiographic finding. Skin testing with purified protein derivative tuberculin was positive with 10-by 7-mm erythema, without an induration or surrounding erythema. There was no family history of a similar reaction to BCG vaccination. The history of the girl's grandmother revealed pulmonary tuberculosis in her twenties. In addition, the possibility of close contact with other tuberculosis patients could not be excluded.We punctured the abscess and submitted the pus for conventional microbial culture, mycobacterial culture, amplification of mycobacterial nucleic acid using an AMPLICOR MTB microwell plate assay kit (Roche Diagnostic Systems, Branchburg, N.J.), and microscopic examination of a smear. Conventional microbial culture failed to detect any significant pathogens. The smear examination was negative, but amplification of nucleic acid using a primer for a Mycobacterium tuberculosis complex-specific region of the 16S rRNA gene (KY172-T3) was positive for M. tuberculosis complexes. A 4-week mycobacterial culture grew an M. tuberculosis complex colony. This colony showed sensitivity to isoniazid, rifampin, ethambutol, and streptomycin and resistance to pyrazinamide, suggesting that the colony may have derived from BCG Tokyo 172. However, we could not clearly distinguish the origin among M. tuberculosis complex strains (M. tuberculosis, M. bovis BCG, or other strains of M. bovis).For the purpose of quick and accurate identification of the sample, we ran multiplex PCR on the sample by a simple modification of the method of Bedwell et al. (1). In 1999, a DNA microarray clarified that 16 regions in BCG strains, named RD1 to RD16, were different from those in M. tuberculosis H37Rv and that some regions differed among BCG strains (2). RD1 is not present in all M. bovis BCG strains, but it is present in other strains of the M. tuberculosis complex. For M. bovis BCG strains, Talbot et al. designed primers to amplify the complement sequence of RD1 (9). Another PCR procedure based on the intergenic region named SenX3-RegX3 was reporte...

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“…However, an important drawback of the current spoligotyping method is its limited discriminatory power compared to that of IS6110 RFLP typing (20,22,42). On the other hand, for M. tuberculosis complex strains containing small numbers of IS6110 copies, including Mycobacterium bovis, spoligotyping is generally more discriminatory than IS6110 RFLP typing (3).…”

mentioning

confidence: 97%

Improvement of Differentiation and Interpretability of Spoligotyping for Mycobacterium tuberculosis Complex Isolates by Introduction of New Spacer Oligonucleotides

Zanden

Kremer²,

Schouls³

et al. 2002

J Clin Microbiol

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The direct repeat (DR) region in Mycobacterium tuberculosis complex strains is composed of multiple well-conserved 36-bp DRs interspersed with nonrepetitive DNA spacer sequences of similar size. Clinical isolates show extensive polymorphism in this DR region, and this has led to the development of a 43-spacer reversed line blot methodology: spoligotyping. Although this method has contributed significantly to the molecular epidemiology of tuberculosis in the last decade, the discriminatory power and the readability of this method were not found to be optimal. In order to improve the discriminatory power, the usefulness of 43 redesigned oligonucleotides and the usefulness of 51 new spacer oligonucleotides were evaluated. For 314 M. tuberculosis complex strains isolated in the central part of The Netherlands over a 5-year period, 264 different IS6110 RFLP types could be distinguished, and 160 different spoligotype patterns were identified by traditional spoligotyping. After the introduction of 51 new spacer oligonucleotides, 14 additional spoligotypes were recognized. This enabled us to split 11 clusters of isolates identified by the traditional spoligotyping. Furthermore, on the basis of the new spacer oligonucleotides a dichotomy was found among the Beijing genotype isolates. Among 76 Mycobacterium bovis strains, 20 patterns were found by traditional spoligotyping and 30 patterns were found by novel probe spoligotyping, respectively. Nine M. bovis subsp. caprae isolates yielded six patterns by traditional spoligotyping and eight patterns by novel probe spoligotyping. A part of the redesigned oligonucleotides slightly improved the reading of spoligotype patterns. The reproducibility of spoligotyping, based on internal control probes, invariably yielded a high score; only 4 (1%) of the 314 patient isolates gave discrepant results. Analysis of a set of 31 duplicate M. tuberculosis complex strains demonstrated a 10% error rate for the identification of blinded duplicate samples. In a redundancy analysis, 40 essential spacer oligonucleotides of the 94-spacer sequences were selected, yielding the same number of spoligotype patterns. We propose to leave the traditional commercialized first-generation membrane for spoligotyping unchanged for current applications and to introduce a second-generation spoligotyping membrane whenever extended discrimination is required, e.g., for low-copy-number IS6110 strains or for phylogenetic studies of Beijing genotype strains.Molecular typing of Mycobacterium tuberculosis complex isolates is a useful tool for epidemiological studies on different levels. In the last decade the application of different DNA fingerprinting techniques has contributed significantly to our understanding of the transmission of tuberculosis (15,45). Actually, these techniques provide a more sensitive means of identifying outbreaks than conventional surveillance (4).A wide variety of DNA fingerprinting techniques has been developed to differentiate between M. tuberculosis complex isolates (11,16,20,29,30), and o...

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Identification of substrains of BCG vaccine using multiplex PCR

Cited by 69 publications

References 15 publications

The History of Tuberculosis and Bacillus Calmette–Guérin Vaccine in Iran

The History of Tuberculosis and Bacillus Calmette–Guérin Vaccine in Iran

Multiplex PCR-Identified Cutaneous Tuberculosis Evoked by Mycobacterium bovis BCG Vaccination in a Healthy Baby

Improvement of Differentiation and Interpretability of Spoligotyping for Mycobacterium tuberculosis Complex Isolates by Introduction of New Spacer Oligonucleotides

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