Detection of circulating Mycobacterium tuberculosis-specific DNA by droplet digital PCR for vaccine evaluation in challenged monkeys and TB diagnosis.
Detection of circulating Mycobacterium tuberculosis-specific DNA by droplet digital PCR for vaccine evaluation in challenged monkeys and TB diagnosis.
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通过液滴数字 PCR 检测循环结核分枝杆菌特异性 DNA，用于受攻击猴子的疫苗评估和结核病诊断
DOI：
10.1038/s41426-018-0076-3
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发表时间：
2018-04-24
影响因子：
13.2
通讯作者：
Zhang XL
中科院分区：
文献类型：
--
作者：
Song N;Tan Y;Zhang L;Luo W;Guan Q;Yan MZ;Zuo R;Liu W;Luo FL;Zhang XL
结核分枝杆菌（M.由于耐多药结核病和人类免疫缺陷病毒（HIV）的合并感染的增加，结核病（TB）正在成为更严重的病原体。迫切需要开发一种有效和灵敏的检测方法，用于疫苗开发、结核病早期诊断和结核病治疗中的细菌载量评估。液滴数字聚合酶链反应（ddPCR）是一种新发展的灵敏的绝对定量核酸浓度的PCR方法。在这里，我们使用ddPCR来定量循环的强毒M。结核特异性CFP 10（10-kDa培养滤液蛋白，Rv 3874）和Rv 1768 DNA拷贝数在来自卡介苗（BCG）接种和/或强毒M. tb H37 Rv攻击的恒河猴。我们发现ddPCR比实时荧光定量PCR（qPCR）更敏感，因为ddPCR的CFP 10检测限为1.2拷贝/μl，而qPCR的CFP 10检测限为15.8拷贝/μl。我们证明，在结核分枝杆菌H37 Rv攻击的恒河猴模型中，ddPCR可以在感染3周后检测到CFP 10和Rv 1768 DNA，并且比qPCR早至少2周。DdPCR还可以成功地定量临床TB患者（活动性肺TB、肺外TB（EPTB）和婴儿TB）血液样本中的CFP 10和Rv 1768 DNA拷贝数。据我们所知，这项研究是第一个证明ddPCR是一种有效和灵敏的方法，用于测量循环CFP 10和Rv 1768 DNA，用于疫苗开发，体内细菌负荷评估和早期TB（包括EPTB和婴儿TB）诊断。
Mycobacterium tuberculosis (M. tb) is emerging as a more serious pathogen due to the increased multidrug-resistant TB and co-infection of human immunodeficiency virus (HIV). The development of an effective and sensitive detection method is urgently needed for bacterial load evaluation in vaccine development, early TB diagnosis, and TB treatment. Droplet digital polymerase chain reaction (ddPCR) is a newly developed sensitive PCR method for the absolute quantification of nucleic acid concentrations. Here, we used ddPCR to quantify the circulating virulent M. tb-specific CFP10 (10-kDa culture filtrate protein, Rv3874) and Rv1768 DNA copy numbers in the blood samples from Bacille Calmette-Guerin (BCG)-vaccinated and/or virulent M. tb H37Rv-challenged rhesus monkeys. We found that ddPCR was more sensitive compared to real-time fluorescence quantitative PCR (qPCR), as the detection limits of CFP10 were 1.2 copies/μl for ddPCR, but 15.8 copies/μl for qPCR. We demonstrated that ddPCR could detect CFP10 and Rv1768 DNA after 3 weeks of infection and at least two weeks earlier than qPCR in M.tb H37Rv-challenged rhesus monkey models. DdPCR could also successfully quantify CFP10 and Rv1768 DNA copy numbers in clinical TB patients' blood samples (active pulmonary TB, extrapulmonary TB (EPTB), and infant TB). To our knowledge, this study is the first to demonstrate that ddPCR is an effective and sensitive method of measuring the circulating CFP10 and Rv1768 DNA for vaccine development, bacterial load evaluation in vivo, and early TB (including EPTB and infant TB) diagnosis as well.
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影响因子：
3.2
作者：
Ganguly, Niladri;Siddiqui, Imran;Sharma, Pawan
通讯作者：
Sharma, Pawan
使用 Xpert MTB/RIF 检测快速有效地诊断结核病和利福平耐药性：荟萃分析
影响因子：
28.2
作者：
Chang, Kai;Lu, Weiping;Chen, Ming
通讯作者：
Chen, Ming
影响因子：
5.5
作者：
Okada, Masaji;Kita, Yoko;Sakatani, Mitsunori
通讯作者：
Sakatani, Mitsunori
DOI：
10.1183/09031936.00214014
发表时间：
2015-04
期刊：
The European respiratory journal
影响因子：
--
作者：
Lönnroth K;Migliori GB;Abubakar I;D'Ambrosio L;de Vries G;Diel R;Douglas P;Falzon D;Gaudreau MA;Goletti D;González Ochoa ER;LoBue P;Matteelli A;Njoo H;Solovic I;Story A;Tayeb T;van der Werf MJ;Weil D;Zellweger JP;Abdel Aziz M;Al Lawati MR;Aliberti S;Arrazola de Oñate W;Barreira D;Bhatia V;Blasi F;Bloom A;Bruchfeld J;Castelli F;Centis R;Chemtob D;Cirillo DM;Colorado A;Dadu A;Dahle UR;De Paoli L;Dias HM;Duarte R;Fattorini L;Gaga M;Getahun H;Glaziou P;Goguadze L;Del Granado M;Haas W;Järvinen A;Kwon GY;Mosca D;Nahid P;Nishikiori N;Noguer I;O'Donnell J;Pace-Asciak A;Pompa MG;Popescu GG;Robalo Cordeiro C;Rønning K;Ruhwald M;Sculier JP;Simunović A;Smith-Palmer A;Sotgiu G;Sulis G;Torres-Duque CA;Umeki K;Uplekar M;van Weezenbeek C;Vasankari T;Vitillo RJ;Voniatis C;Wanlin M;Raviglione MC
通讯作者：
Raviglione MC
影响因子：
8.4
作者：
Elhassan, Mogahid M.;Elmekki, Miskelyemen A.;Hamid, Mohamed E.
通讯作者：
Hamid, Mohamed E.