PCR-荧光探针法检测婴幼儿和儿童人鼻病毒的初步评估

Preliminary evaluation of PCR fluorescence probe method for detection of human rhinovirus in children and infants

目的通过PCR-荧光探针法和Sanger测序法检测呼吸道感染婴幼儿和儿童人鼻病毒(HRV)的感染情况,以评估PCR-荧光探针法的病毒检测能力,并分析HRV载量对患儿临床指标的影响。方法收集2019年5—7月在遵义医科大学第三附属医院住院收治的具有呼吸道感染症状的175例患儿的鼻咽拭子标本,采用PCR-荧光探针法和Sanger测序法检测HRV感染情况并分析HRV载量对婴幼儿和儿童临床指标的影响。结果PCR-荧光探针法检出阳性标本51份(51/175,29.14%),阴性标本124份(124/175,70.86%)。Sanger测序法检出阳性标本48份(48/175,27.43%),阴性标本127份(127/175,72.57%)。与Sanger测序法相比,PCR-荧光探针法检测HRV的灵敏度和特异度分别为100.00%和97.64%,阳性预测值和阴性预测值分别为94.12%和100.00%,一致率为95.80%。两种方法对HRV的检出率比较,差异无统计学意义(P>0.05)。高载量病毒组和低载量病毒组住院天数、入院前最高体温、发热天数、白细胞计数、淋巴细胞百分比及降钙素原比较,差异均无统计学意义(P>0.05),超敏C反应蛋白比较,差异有统计学意义(P<0.05)。结论PCR-荧光探针法检测HRV灵敏度高、特异度高、检测周期短、费用较低且操作简便快捷,可在临床检测HRV中广泛推广。此外,高载量HRV的婴幼儿和儿童的超敏C反应蛋白水平较低载量HRV的婴幼儿和儿童更高。

Objective To evaluate the virus detection ability of PCR fluorescent probe method,and analyze the effect of HRV load on clinical indicators of patients by detecting human rhinovirus(HRV)infection in patients with respiratory tract infection by PCR fluorescent probe method and Sanger sequencing method.Methods The nasopharyngeal swab specimens of totally 175 patients with respiratory tract infection symptoms in the Third Affiliated Hospital of Zunyi Medical University from May to July 2019 were collected,and the PCR-fluorescent probe method and Sanger sequencing method were used to detect HRV infection status and the impact of HRV load on clinical indicators of children and infants was analyzed.Results A total of 51 positive specimens(51/175,29.14%)and 124 negative specimens(124/175,70.86%)were detected by the PCR-fluorescence probe method.A total of 48 positive specimens(48/175,27.43%)and 127 negative specimens(127/175,72.57%)were detected by Sanger sequencing method.Compared with the Sanger sequencing method,the sensitivity and the specificity of PCR-fluorescent probe method for detecting HRV were 100.00%and 97.64%,the positive predictive value and negative predictive value were 94.12%and 100.00%respectively,and the agreement rate was 95.80%.There was no statistically significant difference on the detection rate of HRV by the two methods(P>0.05).There was no statistically significant differences on the days of hospitalization,maximum body temperature before admission,days of fever,white blood cell count,lymphocyte percentage,procalcitonin between high viral load group and low viral load group(P>0.05),while there was statistically significant difference on hypersensitivity C-reactive protein between high viral load group and low viral load group(P<0.05).Conclusion The PCR-fluorescent probe method for detecting HRV has good sensitivity,high specificity,short detection cycle,low cost,simple and quick operation,and could be widely used in clinical detection of HRV.In addition,children and infants with high HRV load have higher levels of hypersensitivity C-reactive protein than children and infants with low HRV load.