Background:
For evaluation of rapid and sensitive tools for diagnosis of tuberculosis, leprosy and HBV infection, and reliable tools for prenatal diagnosis and carrier detection of hemophilia A, we have performed polymerase chain reaction(PCR)
amplifications
using various clinical specimens.
Methods:
PCR amplifications were used to detect tuberculosis, hepatitis B and hemophilia A from various clinical specimens. In the PCR of tuberculosis, oligonucleotide pairs for M. tuberculosis IS 110 DNA were used as primers and m. tuberculosis H37Rv
strain
was used as a positive control, and amplified products of 123 bp fragments were detected. In the PCR of leprosy, oligonucleotide pairs specific for M. leprae DNA were used as primers and amplified products of 530 bp fragments were detected. In
the
PCR
of hepatitis B, oligonucleotide pairs specific for HBV DNA (1763-1892 and 2023-2003) were used as primers and amplified products of 270 bp fragments were detected. In the PCR of Bcl I-RFLP, oligonucleotide pairs for intron 18 in fator VIII gene
were
used as primers and after digestion of 142 bp amplified product with Bcl I, two alleles i.e. negative allele(142 bp) and positive allele (99 and 43 bp) were identified.
Results:
In the M. tubercuosis PCR, positive results were obtained in 16 of 20(80.0%) sputums and seven of 10(70.0%) cerebrospinal fluids. In the M. leprae PCR, positive results were obtained in four of four(100%) biopsy specimens of Lepromatous
leprosy(LL)
and three of three(100%) biopsy specimens of Borderline lepromatous leprosy (BL). In the HBV PCR, positive results were obtained in 40 of 40 (100%) HBs antigen positive sera, and four HBs antigen negative sera were all negative by PCR. In the PCR
of Bcl
I RFLOP within factor VIII, gene frequency with negative allele and positive allele were 15.5% and 84.5% respectively, and female heterozygosity rate was 28.1% and heterozygosity rate of hemophilic carrier was 31.1%.
Conclusions:
These results showed that the PCR provides very sensitive and effective tools for the accurate and rapid diagnosis of tuberculosis, leprosy, hepatitis B and hemophilia A.
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