Relationship of Brucella in blood with clinical symptoms and antibody levels in brucellosis
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摘要:
目的 应用巢式聚合酶链式反应（PCR）方法检测布鲁氏菌病（布病）患者血液中布鲁氏菌DNA, 分析其与临床症状和抗体水平之间的关系。 方法 对门诊就诊患者进行个案调查，包括临床症状、既往史、接触史、用药史等。 布病可疑患者，采集血液，分离血清，进行血清试管凝集试验（SAT），检测布鲁氏菌抗体，按照《布鲁氏菌病诊断》（WS 269—2019）进行布病临床诊断。 同时对患者抗凝全血进行细菌DNA提取，应用巢式PCR方法检测血液中布鲁氏菌DNA。 对不同症状、抗体水平血液中布鲁氏菌细菌数之间的关系进行统计学分析。 结果 118例布病患者中，男性78例，女性40例。 患者的临床症状主要有发热、乏力、头痛、胸背酸痛、腰痛、四肢酸痛、关节肌肉疼痛、出汗、失眠和睾丸痛等。118例布病患者血液标本中，布鲁氏菌DNA阳性率为63.56%（75/118），布鲁氏菌细菌数中位数(MED)为7拷贝/mL血液，四分位数(IQR)为2~17拷贝/mL血液。 118例布病患者中，SAT检测阳性率为38.14%（45/118）。118例布病患者中，新发病例和复诊病例分别占55.08%（65/118）和44.92%（53/118），新发病例和复诊病例血液中布鲁氏菌DNA阳性率分别为72.31%（47/65）和52.83%（28/53）。47例新发病例血液标本中布鲁氏菌细菌数MED为3拷贝/mL血液，IQR为2～26拷贝/mL血液；28例复诊病例血液标本中布鲁氏菌细菌数MED为4拷贝/mL血液， IQR为2～12拷贝/mL血液。 结论 巢式PCR方法检测布病疑似患者血液中布鲁氏菌DNA，有助于布鲁氏菌感染或血清抗体阴性患者的诊断，血液中布鲁氏菌DNA的检测和血清学检查相结合可以有效地诊断布病。 Abstract:Objective To detect Brucella genomic DNA copy number in blood samples from brucellosis by a nested-polymerase chain reaction assay (nested-PCR). The relationship of Brucella DNA copy number with antibody level and clinical symptoms in brucellosis were analyzed. Methods The suspected brucellosis in outpatient department were investigated, the information about their clinical symptoms, exposure and medical history were recorded, and blood samples were collected from them. The Brucella antibody were detected by serum agglutination test (SAT). Brucellosis was diagnosed according to “Brucellosis Diagnosis Criteria” (WS 269—2019). Bacterial genomic DNA was extracted. The genomic DNA of Brucella was detected by nested-PCR assay. The relationship of Brucella DNA copy number with antibody levels and clinical symptoms of brucellosis were analyzed by statistical methods. Results In 118 confirmed brucellosis cases, 78 were men, 40 were women. The clinical symptoms included fever, fatigue, headache, chest and back pain, low back pain, limb pain, joint-muscle pain, sweating and testicular pain. In the 118 cases of brucellosis, the Brucella DNA positive rate was 63.56% (75/118) in the anticoagulant whole blood. The median (MED) of Brucella DNA copy numbers were 7 copies/mL in blood, and the interquartile range (IQR) was 2−17 copies/mL in blood. The positive rate of SAT was 38.14% (45/118) in the 118 cases. The new cases and revisited cases accounted for 55.08% (65/118)and 44.92% (53/118) respectively in 118 cases. The positive rates of Brucella DNA were 72.31% (47/65) and 52.83% (28/53) in new cases and revisited cases respectively. The MED of Brucella DNA copy numbers were 3 copies/mL in blood, the IQR was 2−26 in 47 new cases. The MED of Brucella DNA copy numners were 4 copies/mL in blood, the IQR was 2−12 copies/mL in blood in 28 revisited cases. Conclusion In suspected brucellosis patients, attention should be paid to those with joint-muscle pain. The PCR detection was helpful in the diagnosis of early infection of Brucella or brucellosis patients with negative SAT results. The combination of Brucella DNA detection and serological examination could facilitate the diagnosis of brucellosis. -
Key words:
- Brucellosis /
- Symptom /
- Brucella /
- Brucella antibody /
- Nested-polymerase chain reaction assay
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表 1 118例布病患者临床特征与血液标本中布鲁氏菌检测结果分析
Table 1. Analysis on clinical characteristics and Brucella detection results of 118 brucellosis
特征 检测数
（例）阳性数
（例）阳性率
（%）χ2值 P值 性别 0.029 0.865 男性 78 50 64.10 女性 40 25 62.50 年龄组（岁） 0.255 0.614 ≤45 53 35 66.04 >45 65 40 61.54 发热 0.055 0.815 无 100 64 64.00 有 18 11 61.11 乏力 0.015 0.903 无 94 60 63.83 有 24 15 62.50 头痛 0.132 0.716 无 111 71 63.96 有 7 4 57.14 胸背疼痛 0.595 0.440 无 105 68 64.76 有 13 7 53.85 腰痛 3.581 0.058 无 98 66 67.35 有 20 9 45.00 四肢酸痛 0.015 0.903 无 76 48 63.16 有 42 27 64.29 关节疼痛 2.147 0.143 无 22 11 50.00 有 96 64 66.67 出汗 1.686 0.194 无 100 66 66.00 有 18 9 55.00 血清试管凝集试验 0.303 0.582 阳性 45 30 66.67 阴性 73 45 61.64 合计 118 75 63.56 
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表 2 新发病例与复诊病例血液标本布鲁氏菌检出结果
Table 2. Brucella detection results of new cases and follow-up cases
病例类型 检测数（例） 阳性数（例） 阳性率（%） χ2值 P值 细菌计量（拷贝/mL血液） 中位数 四分位数 P值 新发病例 65 47 72.31 4.782 0.029 3 2～26 2.180 复诊病例 53 28 52.83 4 2～12 合计 118 75 63.56
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