Genotyping by multiplex polymerase chain reaction for detection of endemic hepatitis B virus transmission.
A nested polymerase chain reaction (PCR) protocol was developed for rapid genotyping of hepatitis B virus (HBV). During the first PCR round, a universal HBV primer pair was used to amplify the entire pre-S region of the HBV genome. Within the pre-S region, many nucleotide exchanges are observed. These are partly correlated to the serological hepatitis B surface antigen subtypes. Five additional subtype-specific primers were selected from that region which, together with two universal non-group-specific primers, generated specific combinations of two to four DNA fragments of defined sizes. By this approach, 55 hepatitis B surface antigen-positive patients from a pediatric oncology unit in Germany were analyzed. Fifty-four patients who had been infected within 2 years had an identical pattern in the multiplex PCR, suggesting a common source of infection and person-to-person transmission within the unit. One child who was infected 5 years later had a different PCR pattern and, therefore, must have been infected from a different source. Furthermore, 109 serum samples taken from pregnant Cameroonian women and 25 serum samples from their babies taken 6 months after birth were analyzed. In one case, mother-to-infant transmission of the virus was demonstrated. Apart from its role in epidemiological studies on HBV, multiplex PCR may also be a useful tool for rapid genetic analysis in other fields if there is a moderate degree of sequence variation which enables the design of specific primers.
Department of Pediatrics, Justus-Liebig-University, Giessen, Germany.
R
R
Repp
S
S
Rhiel
K H
KH
Heermann
S
S
Schaefer
C
C
Keller
P
P
Ndumbe
F
F
Lampert
W H
WH
Gerlich
D001483:Base Sequence; D002648:Child; D003428:Cross Infection / Q000453:epidemiology / Q000382:microbiology; D015342:DNA Probes; D004279:DNA, Viral / Q000235:genetics; D004196:Disease Outbreaks; D005260:Female; D005838:Genotype; D005858:Germany / Q000453:epidemiology; D006509:Hepatitis B / Q000453:epidemiology / Q000382:microbiology / Q000635:transmission*; D006515:Hepatitis B virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D007223:Infant; D008431:Maternal-Fetal Exchange; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy
D015342:DNA Probes; D004279:DNA, Viral