Department of Infectious Diseases, Rigshospitalet, Copenhagen, Denmark.
Clin Chem Lab Med. 2011 Jul;49(7):1171-5. doi: 10.1515/CCLM.2011.184. Epub 2011 Apr 20.
Understanding the distribution of antiretro-virals in breastfeeding HIV-positive mothers is essential, both for prevention of mother-to-child HIV transmission and for research on the development of drug resistance. The ARK nevirapine (NVP)-test is an immunoassay method for nevirapine measurements, developed and validated for plasma use. In this study, the ARK NVP-test was evaluated for measurement of nevirapine concentrations in breast milk. High performance liquid chromatography (HPLC) is the method currently used to determine nevirapine in breast milk. This method, however, requires complicated extraction techniques. The ARK method employs an immunoassay technology and requires a small sample volume (40 μL) and no pre-treatment of the samples.
Commercial enzyme and antibody were used and calibration standards and quality controls were prepared from pooled breast milk from HIV-uninfected women. Clinical samples from HIV-infected women receiving a single-dose of nevirapine were analyzed.
Precision and accuracy were evaluated with two concentrations of quality control materials analyzed in three replicates on four different days and was <4%, and between 96.5% and 104.6%, respectively. Clinical samples were analyzed and CVs ranged from 0.0% to 11.1%. The median nevirapine concentration in breast milk 1 week post-partum was 0.29 μg/mL (range 0.11-0.90 μg/mL) in women treated with a single-dose of nevirapine.
The ease of use and small sample volume makes the ARK assay an attractive alternative to HPLC analyses for determinations of nevirapine concentrations in breast milk.
了解母乳喂养的 HIV 阳性母亲体内抗逆转录病毒的分布情况对于预防母婴传播 HIV 和研究药物耐药性的发展至关重要。ARK 奈韦拉平（NVP）检测是一种用于检测奈韦拉平的免疫分析法，已开发并验证可用于血浆检测。本研究评估了 ARK NVP 检测法用于检测母乳中奈韦拉平浓度的能力。高效液相色谱法（HPLC）是目前用于检测母乳中奈韦拉平的方法，但需要复杂的提取技术。ARK 方法采用免疫分析法技术，仅需小样本量（40μL），且无需对样本进行预处理。
使用商业酶和抗体，从未感染 HIV 的女性的混合母乳中制备校准标准和质控品。分析接受单次奈韦拉平剂量的 HIV 感染女性的临床样本。
使用两种浓度的质控材料在四个不同日期的三个重复中进行评估，精密度和准确度均<4%，分别为 96.5%至 104.6%。分析临床样本时，CV 范围为 0.0%至 11.1%。产后 1 周时，单次奈韦拉平剂量治疗的女性母乳中奈韦拉平的中位数浓度为 0.29μg/mL（范围为 0.11-0.90μg/mL）。
由于使用方便且样本量小，ARK 检测法成为 HPLC 分析检测母乳中奈韦拉平浓度的一种有吸引力的替代方法。
