Comparative Genomic Analysis of Haemophilus haemolyticus and Nontypeable Haemophilus influenzae and a New Testing Scheme for Their Discrimination
Comparative Genomic Analysis of Haemophilus haemolyticus and Nontypeable Haemophilus influenzae and a New Testing Scheme for Their Discrimination
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DOI：
10.1128/jcm.01511-16
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发表时间：
2016-12-01
影响因子：
9.4
通讯作者：
Wang, Xin
中科院分区：
文献类型：
--
作者：
Hu, Fang;Rishishwar, Lavanya;Wang, Xin
最近发现溶血性嗜血杆菌有可能导致侵袭性疾病。它与非分型流感嗜血杆菌(NT H.Inflenzae)密切相关。NT型流感嗜血杆菌和溶血性嗜血杆菌经常被误诊，因为现有针对已知的溶血性嗜血杆菌表型的检测都不能特异性地鉴定溶血性嗜血杆菌。通过对溶血性嗜血杆菌和流感嗜血杆菌的比较基因组分析，我们确定了溶血性嗜血杆菌特有的基因，可以作为鉴定溶血性嗜血杆菌的靶标。建立了一种针对编码嘌呤合成途径中的磷酸核糖基甘氨酰胺甲酰转移酶2的实时定量聚合酶链式反应方法。该方法的检测下限为40个基因组/聚合酶链式反应，检测溶血性嗜血杆菌的灵敏度为98.9%，特异度为97%。为了提高对溶血性嗜血杆菌和流感嗜血杆菌的鉴别能力，还对两种检测方法(溶血性嗜血杆菌和编码蛋白D脂蛋白的流感嗜血杆菌Hpd)联合检测方案进行了评价，对溶血性嗜血杆菌和流感嗜血杆菌的敏感性分别为96.7%和98.2%，对流感嗜血杆菌的敏感性和特异性分别为92.8%和100%。该双靶点检测方案可用于诊断和监测由溶血性嗜血杆菌和流感嗜血杆菌引起的感染和疾病。
Haemophilus haemolyticus has been recently discovered to have the potential to cause invasive disease. It is closely related to nontypeable Haemophilus influenzae (NT H. influenzae). NT H. influenzae and H. haemolyticus are often misidentified because none of the existing tests targeting the known phenotypes of H. haemolyticus are able to specifically identify H. haemolyticus. Through comparative genomic analysis of H. haemolyticus and NT H. influenzae, we identified genes unique to H. haemolyticus that can be used as targets for the identification of H. haemolyticus. A real-time PCR targeting purT (encoding phosphoribosylglycinamide formyltransferase 2 in the purine synthesis pathway) was developed and evaluated. The lower limit of detection was 40 genomes/PCR; the sensitivity and specificity in detecting H. haemolyticus were 98.9% and 97%, respectively. To improve the discrimination of H. haemolyticus and NT H. influenzae, a testing scheme combining two targets (H. haemolyticus purT and H. influenzae hpd, encoding protein D lipoprotein) was also evaluated and showed 96.7% sensitivity and 98.2% specificity for the identification of H. haemolyticus and 92.8% sensitivity and 100% specificity for the identification of H. influenzae, respectively. The dual-target testing scheme can be used for the diagnosis and surveillance of infection and disease caused by H. haemolyticus and NT H. influenzae.