A two-stage polymerase chain reaction (PCR) assay employing oligonucleotide primers from the B1 gene of Toxoplasma gondii was developed and assessed for sensitivity and specificity. It was able to detect T. gondii DNA from as little as one parasite/sample in mock-infected rat or mouse leucocyte preparations. Parasitaemia was also identified in animals at five stages between 16 and 66 h after infection with the virulent RH strain, and at 12 stages between 2 and 38 days after infection with the cyst-forming Beverley strain. In the latter case, PCR was more sensitive than animal culture. No cross-reactions were observed in samples containing various opportunist pathogens which may also be found in the blood of immunocompromised patients.
LuftBJ,
RemingtonJS.
Toxoplasmosis of the central nervous system. In:
RemingtonJS.
(ed) Current clinical topics in infectious disease New York: McGraw-Hill; 1985; 6315–358
DaifosF,
ForestierF,
Cappela-PavlovskyM.
et al. Prenatal management of 746 pregnancies at risk for congenital toxoplasmosis. N Engl J Med1988; 318:271–275
HollimanRE,
JohnsonJD,
SavvaD.
Diagnosis of cerebral toxoplasmosis in association with AIDS using the polymerase chain reaction. Scand J Infect Dis1990; 22:243–244
BurgJL,
GroverCM,
PoulettyP,
BoothroydJC.
Direct and sensitive detection of a pathogenic protozoan, Toxoplasma gondii, by polymerase chain reaction. J Clin Microbiol1989; 27:1787–1792
van KnapenF,
PanggabeanSO,
van LeusdenJ.
Demonstration of toxoplasma antigen containing complexes in active toxoplasmosis. J Clin Microbiol1985; 22:645–650
HasslA,
AspöckH,
FlammH.
Circulating antigen of Toxoplasma gondii in patients with AIDS: significance of detection and structural properties. Zentralbl Bakteriol Mikrobiol Hyg1988; A270:302–309
WeissLM,
UdemSA,
SalgoM,
TanowitzHB,
WittnerM.
Sensitive and specific detection of toxoplasma DNA in an experimental murine model: use of Toxoplasma gondii-specific cDNA and polymerase chain reaction. J Infect Dis1991; 163:180–186
WagesJM,
HamdallahM,
CalabroMA.
et al. Clinical per formance of a polymerase chain reaction testing algorithm for diagnosis of HIV-1 infection in peripheral blood mononuclear cells. J Med Virol1991; 33:58–63
BeverleyJKA.
Some aspects of toxoplasmosis, a world wide zoonosis. In:
SoulsbyEJL.
(ed) Parasitic zoonoses: clinical and experimental studies New York: Academic Press; 19741–25
BronsteinI,
VoytaJC,
LazzariKG,
MurphyO,
EdwardsB,
KrickaLJ.
Rapid and sensitive detection of DNA in southern blots with chemiluminescence. Biotechniques1990; 8:310–314
Ho-YenDO,
JossAWL,
BalfourAH,
SmythETM,
BairdD,
ChattertonJMW.
An assessment of Toxoplasma gondii parasitaemia by PCR in human blood samples. J Clin Pathol1992 in press