Stochastics and Biology Group, Probability and Statistics (LPSM, CNRS 8001), Sorbonne University, Campus Pierre et Marie Curie, 4 Place Jussieu, 75005, Paris, France.
MERIT (IRD 261), University of Paris, Faculté de Pharmacie, 4 Avenue de l'Observatoire, 75006, Paris, France.
Malar J. 2021 Jan 5;20(1):6. doi: 10.1186/s12936-020-03530-z.
BACKGROUND: Despite many technological advances for malaria parasite detection (e.g. high resolution image acquisition), microscopic reading of thick blood smear (TBS) remains the gold standard. Even though available in low technology environment, the microscopy of TBS is slow and time consuming. Moreover microscopy may induce errors at many levels and has no quality control. METHODS: A electronic extension of the mechanical tally counter is proposed. In addition to the counting process it includes the process of counting itself that relies on the time elapsed between two successive pressures of the counting button leading to a timed tally counter (TTC). The microscopist performs the reading with the specific instruction starting by counting, in each high power fields, leucocytes first and then parasites. The time-stamp of all pressures of counting buttons are recorded along with the nature of the count. The data are recorded internally in CSV format and are exportable. The detection of HPFs locations and leukocyte/parasite counts per HPFs is performed through a hidden semi-Markov model (with outliers) allowing both to take into account the known distribution of leukocyte per HPFs (using a negative binomial distribution) and the pauses and hesitation of the microscopist during the reading. Parameters are estimated via the expectation-maximization algorithm. Hyper-parameters are calibrated using expert annotations. Forward/backward recursions are used to obtain the HPFs locations. RESULTS: This approach provides richer data at no extra cost. It has been demonstrated that the method can derive parasites per HPF, leukocytes per HPF, and parasite/leukocyte ratio with robust non-parametric confidence intervals. Moreover a direct digital data entry leads to a less expensive process and decreased time-consuming and error-prone manual data entry. Lastly the TTC allows detecting possible protocol break during reading and prevents the risk of fraud. DISCUSSION AND CONCLUSION: Introducing a programmed digital device in the data acquisition of TBS reading gives the opportunity to develop easily new (possible adaptive) reading protocols that will be easily followed by the reader since they will be embedded directly in the device. With the TTC the reader only has to read HPFs, counting leukocytes first and parasites second, and the counter will beep when the protocol is completed.
背景:尽管疟疾寄生虫检测技术有了许多进步(例如高分辨率图像采集),但厚血涂片(TBS)的显微镜检查仍然是金标准。尽管在低技术环境下可用,但 TBS 的显微镜检查既缓慢又耗时。此外,显微镜检查可能会在多个层面引起误差,并且没有质量控制。
方法:提出了机械计数器的电子扩展。除了计数过程外,它还包括计数本身的过程,该过程依赖于连续按下计数按钮之间的时间间隔,从而形成定时计数器(TTC)。显微镜检查师按照特定说明进行阅读,首先在每个高倍视野中计数白细胞,然后计数寄生虫。记录所有计数按钮压力的时间戳以及计数的性质。数据以 CSV 格式内部记录,并可导出。通过隐藏的半马尔可夫模型(带异常值)检测 HPF 位置和每个 HPF 的白细胞/寄生虫计数,该模型既可以考虑已知的每个 HPF 白细胞分布(使用负二项分布),也可以考虑显微镜检查师在阅读过程中的停顿和犹豫。通过期望最大化算法估计参数。超参数使用专家注释进行校准。使用前向/后向递归获得 HPF 位置。
结果:这种方法以零额外成本提供了更丰富的数据。已经证明,该方法可以以稳健的非参数置信区间得出每个 HPF 的寄生虫、每个 HPF 的白细胞和寄生虫/白细胞比值。此外,直接数字数据输入可降低流程成本,减少耗时且易出错的手动数据输入。最后,TTC 可以检测到阅读过程中可能出现的协议中断,并防止欺诈风险。
讨论与结论:在 TBS 阅读数据采集过程中引入编程数字设备,可以轻松开发新的(可能是自适应的)阅读协议,由于这些协议直接嵌入到设备中,因此读者可以轻松遵循。使用 TTC,读者只需阅读 HPF,首先计数白细胞,然后计数寄生虫,当协议完成时计数器会发出蜂鸣声。
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