Massi M Nasrum, Shirakawa Toshiro, Gotoh Akinobu, Bishnu Acharya, Hatta Mochammad, Kawabata Masato
International Center for Medical Research, Kobe University School of Medicine, Kobe, Japan.
Int J Med Microbiol. 2005 Jun;295(2):117-20. doi: 10.1016/j.ijmm.2005.01.003.
We quantified the gene copies from Salmonella enterica serovar Typhi (S. Typhi) in the blood of patients suspected of having typhoid fever by using TaqMan-based real-time PCR (TaqMan assay) to target the S. Typhi flagellin gene in genomic DNAs isolated from blood samples. Of 55 blood samples taken from suspected typhoid fever patients, eight blood samples with a positive blood culture had S. Typhi loads ranging from 1.01 x 10(3) to 4.35 x 10(4) copies/ml blood, and from 47 blood samples with negative blood culture, there were 40 (85.1%) TaqMan assay-positive samples with loads ranging from 3.9 to 9.9 x 10(2) copies/ml blood. In the present study, the TaqMan assay detected more than 10(3) copies/ml blood of S. Typhi in all of the blood culture-positive samples, whereas less than 10(3) copies/ml blood of S. Typhi were detected in the blood culture-negative samples. Our findings suggest that a TaqMan assay may be useful for assessing S. Typhi loads, especially in cases of suspected typhoid fever with negative results from the standard blood culture test.
我们通过基于TaqMan的实时聚合酶链反应（TaqMan检测法），以从血样中分离出的基因组DNA中的伤寒杆菌鞭毛蛋白基因为靶点，对疑似伤寒热患者血液中的伤寒杆菌血清型伤寒杆菌（伤寒杆菌）基因拷贝进行了定量分析。在从疑似伤寒热患者采集的55份血样中，8份血培养呈阳性的血样中伤寒杆菌载量为每毫升血液1.01×10³至4.35×10⁴拷贝，而在47份血培养呈阴性的血样中，有40份（85.1%）TaqMan检测呈阳性，载量为每毫升血液3.9至9.9×10²拷贝。在本研究中，TaqMan检测法在所有血培养阳性样本中检测到每毫升血液中伤寒杆菌超过10³拷贝，而在血培养阴性样本中检测到每毫升血液中伤寒杆菌少于10³拷贝。我们的研究结果表明，TaqMan检测法可能有助于评估伤寒杆菌载量，尤其是在标准血培养检测结果为阴性的疑似伤寒热病例中。
