Development and Evaluation of a New Measles Detection Assay Using Real-Time RT-PCR

被引:0
作者
Chayeb, Vera A. [1 ]
Dolgova, Anna S. [1 ]
Popova, Margarita R. [1 ]
Zheleznova, Nina V. [1 ]
Shirobokova, Svetlana A. [1 ]
Shabalina, Anna V. [1 ]
Sharova, Alena A. [1 ]
Gladkikh, Anna S. [1 ]
Antipova, Anastasia Yu. [1 ]
Kirichenko, Anastasiia D. [1 ]
Ramsay, Edward S. [1 ]
Dedkov, Vladimir G. [1 ,2 ]
机构
[1] St Petersburg Pasteur Inst, Fed Serv ConsumersRights Protect & Human Well Bein, St Petersburg 197101, Russia
[2] First Moscow State Med Univ Sechenov Univ, Martsinovsky Inst Med Parasitol Trop & Vector Born, Moscow 119048, Russia
关键词
measles detection; real-time RT-PCR; molecular methods; outbreak; RISK-FACTORS; VIRUS; INFECTION; OUTBREAK; CHILDREN;
D O I
10.3390/ijms26051801
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The severity of MeV infection has been greatly reduced by the development of a live attenuated vaccine, which has been incorporated into vaccination programs in many countries. However, poor access to health facilities, and above all, the increase in anti-vaccination movements, has prevented the achievement of sufficient vaccination coverage. In outbreak scenarios, a rapid and transportable method can improve differential diagnosis, including removing ambiguity in suspected measles cases, contacts, or a cohort. In response to the need, we have developed a new RT-qPCR-based MeV detection assay. The LOD of the developed assay was determined on different PCR machines and the higher threshold was 1-1.2 103 copies/mL. The joint diagnostic sensitivity of ELISA and RT-PCR (used together) was 100%, and used combinedly, these two methods enable detection of all measles-infected persons, which is extremely important for controlling contagion and spread of infection. During the clinical validation of the assay on 200 clinical samples from measles-suspected cases using ELISA, 157 samples showed a positive result, while 163 positive cases were confirmed by the RT-qPCR assay. The concordance between the two techniques was 93%. According to our results, the real-time RT-PCR approach used in our study is more sensitive and appears to be a more promising method for measles diagnosis during early stages of the disease, likely before the rise of specific IgM antibodies detected by ELISA.
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页数:13
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