ABSTRACT
The aim of this study is to establish a multiplex polymerase chain raction (PCR) to identify of four kinds of laboratory animal pathogens: Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae and Klebsiella pneumoniae.Methods Specific primers were designed based on GenBank data.The multiplex PCR system was established through optimization of multiple PCR and detection of its specificity and sensitivity.This technique was used to test artificially infected samples and tracheal secretions of experimental animals (rat, mouse, guinea pig, rabbit, hamster), and comparing the detection results by this method and traditional detection test.Results Target bands of Pasteurella multocida (356 bp), Bordetella bronchiseptica (237 bp), Mycoplasma pneumoniae (266 bp), and Klebsiella pneumoniae (142 bp) were obtained, with a detection sensitivity of Klebsiella pneumoniae of 10 pg, and that of Pasteurella multocida, Bordetella bronchiseptica and Mycoplasma pneumoniae of 1 pg by this newly developed multiplex PCR assay.No target bands were observed from the non-specific pathogens of artificially infected samples.The tracheal secretions taken from 45 experimental animals (mice and rabbits) were tested with this new PCR assay, among which 15 cases of Klebsiella pneumonia and 9 cases of Pasteurella multocida were detected as positive, while all the results of traditional method and serological test were negative.Conclusions A simple, rapid, specific and highly sensitive multiplex PCR system has been successfully established.It is valuable for detection of Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae, and Klebsiella pneumoniae in laboratory animals.
ABSTRACT
Objective To discuss the test efficiency of three methods for detecting Toxoplasma IgG antibody. Methods To-tally 304 specimens were detected parallelly for Toxoplasma IgG antibody by using the gold marked method,indirect hemagglutina-tion test(IHA),and enzyme-linked immunosorbent assay(ELISA),and the sensitivity,specificity and Youden index of these methods were compared. Results The detection sensitivities of gold marked method,IHA,and ELISA for Toxoplasma IgG anti-body were 85.5%,89.8%and 91.9%respectively(χ2=4.12,P>0.05);the specificities were 92.4%,96.6%and 97.5%respec-tively(χ2=4.06,P>0.05). The detection efficiency and Youden index of ELISA were 94.1%and 0.89 respectively,being high-er than those of IHA and gold marked method. Conclusion The sensitivity and specificity of the ELISA method for Toxoplasma IgG antibody are higher,and in addition,it can be automated. Therefore,it is suitable for large-scale Toxoplasma IgG antibody screening.
ABSTRACT
Objective To explore the impact of Toxoplasma gondii infection on pregnancy outcomes in early pregnancy wom-en. Methods Toxoplasma gondii IgM and IgG antibodies in the peripheral blood of 2 993 early pregnant women were detected by using enzyme-linked immunosorbent assay(ELISA). According to the test results,the infected ones were divided into an acute in-fection group,a previous infection group,and an active infection group,and 200 pregnant women without Toxoplasma infection were randomly chosen as a control group,and the pregnancy outcomes of the four groups were followed up and the results were compared. Results There were 286 women infected with Toxoplasma gondii,with the infection rate of 9.56%(286/2 993),in which 43 cases were diagnosed as acute infection,156 were previous cases,and the other 87 were active infection ones. The inci-dences of adverse pregnancy outcomes in the above 3 groups and the control group were 13.95%(6/43),1.92%(3/156),5.75%(5/87)and 1.50%(3/200),respectively. The incidences of adverse pregnancy outcomes in the acute infection group and active in-fection group were both higher than that in the control group,the differences were statistically significant(both P0.05). Conclusion Acute and ac-tive Toxoplasma gondii infections are closely associated with the occurrence of adverse pregnancy outcomes in early pregnant wom-en;therefore,Toxoplasma gondii IgM antibody should be included in the routine inspection items of the pre-pregnancy physical examination for child-bearing age women.