Seroprevalence and genetic characterization of Toxoplasma gondii in cancer patients in Anhui Province, Eastern China.

BACKGROUND: Recent studies have indicated the predominance of Toxoplasma gondii genotype Chinese 1 in animals in China. However, little is known of the genetic features of the parasite in humans. This study aims to determine the prevalence of anti-T. gondii antibodies based on which the genetic character of the parasite was identified in cancer patients in China. METHODS: A total of 1014 serum samples with malignant neoplasms were collected from six tertiary-care hospitals (HAUCM, APH, HAMU, XAH, FHH and HBMC) from January, 2012 to August, 2013. Antibodies against T. gondii were examined by enzyme-linked immunosorbent assay (ELISA). Blood samples were subsequently used for PCR assay to detect T. gondii DNA (gra6). The DNA positive samples were subjected to genotyping using a multiplex multilocus nested PCR-RFLP at 10 loci, including sag1, sag2, sag3, btub, gra6, l358, c22-8, c29-2, pk1 and apico. Samples from the patients were anonymous and only data with regard to age and gender was available at sample collection. RESULTS: Overall, 8.38% (85/1014) of the examined patients showed positive antibodies against T. gondii. Among them, 61 (6.02%) were seropositive only for IgG, 16 (1.58%) were only for IgM, and 8 (0.79%) were found to be positive for both IgG and IgM. The seroprevalence of antibodies to Toxoplasma ranged from 5.8% to 11.0%, without regional difference (χ(2) = 4.764, P = 0.445). No significant differences of the positive rates of T. gondii infection were noted in genders (male, 8.96%; female, 7.45%) (χ(2) = 0.707, P = 0.400) and in ages (χ(2) = 1.172, P = 0.947). Of 1014 DNA samples, 36 (3.55%) were positive for T. gondii by nested PCR at gra6 locus and nine gave rise to complete genotyping results. All samples with achieved PCR-RFLP genotyping showed a common genetic character of type Chinese 1 (ToxoDB#9). CONCLUSION: Seroprevalence of toxoplasmosis in immunosuppressed individuals is rarely reported in China and we presented a positive rate of 8.38% in cancer patients. Toxoplasma genomic DNA genotyping demonstrated a common genetic character of Chinese 1, indicating a possible pathogenic origin of animals in human infection.

Toxoplasma gondii prevalence in food animals and rodents in different regions of China: isolation, genotyping and mouse pathogenicity.

BACKGROUND: Recent studies of Toxoplasma gondii isolates from animals in different regions of China have shown a limited genetic diversity and type China 1 was the dominant genotype of T. gondii prevalent in Chinese animals. However, little has been known concerning the isolation and genotyping of T. gondii circulating in chickens, pigs and rodents in China. The aim of the study was to characterize samples of T. gondii isolates obtained from naturally infected cats, pigs and free-range chickens slaughtered for human consumption in China. METHODS: In the present study, brain tissues of 77 animals collected from different areas of China, including 24 free-range chickens (Gallus domesticus) , 13 voles (Rattus flavipectus), 23 pigs and 17 cats, were bioassayed in mice and viable T. gondii were isolated from the brains of eleven. These eleven T. gondii isolates were maintained in Kunming (KM) outbred mice and DNA isolated from tissues of infected mice was characterized using 11 PCR-restriction fragment length polymorphism (PCR-RFLP) markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3. Moreover, to determine mouse virulence of China 1 lineage of parasites, a TgCtgy5 genotype isolate was selected randomly and assessed in KM mice with different inoculation doses. RESULTS: Results of genotyping revealed that ten isolates were type China 1 (ToxoDB PCR-RFLP genotype #9), and TgCksz1 was a new genotype that was reported for the first time designated here as ToxoDB PCR-RFLP #225. No clonal types I, II and III lineages were found. DNA sequencing of four introns (EF1, HP2, UPRT1 and UPRT7) and two genes (GRA6 and GRA7) from representative isolates confirmed the results of PCR-RFLP genotyping. The TgCtgy5 isolate was highly virulent in KM mice; all infected mice died of acute toxoplasmosis, irrespective of the inoculation dose. The results indicate that mouse virulent isolates of T. gondii are predominantly circulating in cats in China. CONCLUSIONS: T. gondii isolated from chickens, pigs, cats and rodents in different locations in China were genotyped and the results reconfirmed the limited diversity of T. gondii in China and showed that type China 1 lineage was dominant in this country.