Clinical utility of hyperglycosylated hCG in serum taken before hydatidiform mole evacuation to predict persistent trophoblastic disease

OBJECTIVE: Human chorionic gonadotropin (hCG) is widely used in the management of hydatidiform mole and persistent trophoblastic disease (PTD). Studies on hyperglycosylated human chorionic gonadotropin (invasive trophoblast antigen, ITA) in PTD are limited. In serum samples taken before evacuation of molar pregnancies we measured the concentrations of free hCG beta-subunit (free hCGbeta), "total" hCG (hCG+hCGbeta) and ITA, and determined whether ITA, the two other hCG analytes, or the calculated ratios of hCGbeta/hCG+hCGbeta, hCGbeta/ITA and hCG+hCGbeta/ITA could predict the later development of PTD. DESIGN: A retrospective study based on blood specimens collected in the Dutch Central Registry for Hydatidiform Moles. The study group comprised 97 patients with hydatidiform moles who did not develop PTD after mole evacuation and 33 patients who did develop PTD. Methods: Serum samples from 130 patients with hydatidiform mole with or without PTD were assayed using specific (radio)immunoassays for free hCGbeta, total hCG, and ITA. From these analytes we also calculated the ratios hCGbeta/hCG+hCGbeta, hCGbeta/ITA, and hCG+hCGbeta/ITA. To predict the development of PTD from these analytes and parameters we performed receiver-operating characteristic (ROC) curve analysis, resulting in areas under the curve (AUCs) that represented the diagnostic accuracy which was rated in a range from excellent (AUC >0.9 or <0.1) to poor (AUC 0.4-0.6). Results: The diagnostic accuracy of ITA was moderate (0.618) and not different from that of free hCGbeta (0.610) and hCG+hCGbeta (0.622). CONCLUSIONS: ITA as well as the other analytes and parameters in serum taken prior to evacuation from patients with molar pregnancies cannot be used to predict the subsequent development of persistent trophoblastic disease.     

Prophylactic effect of bestatin on the onset of invasive mole — clinical and fundamental studies

This study was performed to determine whether bestatin (Ubenimex) has clinical prophylactic effects on the onset of invasive mole and a direct inhibitory effect on the growth of hydatidiform molar cells. A total of 49 patients with hydatidiform mole treated at Nagoya University Hospital from 1984 to 1990 were randomly divided into two groups, a bestatin administered-group and a bestatin non-administered group. Patients in the bestatin group were given 30 mg of bestatin orally and daily for three months just after their molar deliveries. There was no significant difference in age, gravidity, parity and gestational weeks between the two groups. There was also no significant difference in the duration of human chorionic gonadotropin (hCG) negative conversion in patients without invasive mole between the two groups. However, the incidence of invasive mole in the bestatin group (2/25, 8%) was significantly lower than that of the non-bestatin group (7/24, 29.2%). Nevertheless, there was no significant difference between the two groups in such immunological parameters as PHA skin test, PPD skin test, PHA stimulation index (PHA-SI), white blood cell (WBC) count lymphocytes % per WBC, OKT 3% per lymphocytes, OKT 4% per lymphocytes, OKT4/OKT8 and Leu 11% per lymphocytes. In vitro studies were performed with primary cultured hydatidiform moles. The result was that bestatin inhibited the secretion of hCG and³H-thymidine uptake of hydatidiform molar cells. Thus, a possibility was suggested that bestatin directly inhibits the growth of hydatidiform molar cells and prevents the onset of invasive mole.     

葡萄胎中MBD2表达及其相关基因甲基化芯片研究

用Real-time RT-PCR、Western blot和免疫组织化学方法分别检测了去甲基化酶MBD2(methyl-CpG-binding domain 2,MBD2)在完全型葡萄胎(complete hydatidiform mole,CHM)和正常早期妊娠绒毛中的表达,用甲基化DNA免疫沉淀MeDIP(methylated DNA immunoprecipitation)-甲基化芯片分析完全型葡萄胎和正常早期妊娠绒毛中相关基因的甲基化情况,用生物信息学分析筛选了差异甲基化基因并进行功能分类。MBD2的mRNA在完全型葡萄胎中的表达明显高于正常早期妊娠绒毛(P=0.0083),Western blot(P=0.0005)和免疫组织化学(P=0.0091)检测到MBD2蛋白表达与Real-time RT-PCR结果一致。结果显示MBD2在完全型葡萄胎中的表达显著高于正常早期妊娠绒毛组织(P<0.01),与正常早期妊娠绒毛组织相比较,完全型葡萄胎组织中相对有89个基因发生了去甲基化,其中85个基因可被映射到基因组图谱中,MBD2在完全型葡萄胎中的高表达及部分基因的去甲基化可能在完全型葡萄胎的发生中扮演了重...     

Comparative study of the mucin-type sugar chains of human chorionic gonadotropin present in the urine of patients with trophoblastic diseases and healthy pregnant women

Human chorionic gonadotropins (hCGs) highly purified from the urine of patients with trophoblastic diseases and of healthy pregnant women contain approximately four mucin-type sugar chains in one molecule. The structures of these sugar chains were studied comparatively by using a new sensitive method to obtain mucin-type sugar chains quantitatively as radioactive oligosaccharides from a small amount of glycoproteins. The mucin-type sugar chains of all hCGs include sialylated and nonsialylated Gal beta 1----3GalNAc and Gal beta 1----4GlcNAc beta 1----6(Gal beta 1----3)GalNAc. In the case of normal hCG and hydatidiform mole hCG, oligosaccharides containing the tetrasaccharide core occupy approximately 10% of the total mucin-type sugar chains. The ratio of the tetrasaccharide containing oligosaccharides is increased prominently to approximately 60% in choriocarcinoma hCG. The proportion in invasive mole hCG was also increased, but less than the proportion of choriocarcinoma hCG.     

Human chorionic gonadotropin promotes thyroid growth via thyrotropin receptors in FRTL-5 cells

To ascertain the presence of thyroid growth-promoting activity (TGA) in the sera of pregnant women, we measured TGA in the sera of pregnant women by means of a bioassay based on [3H]-thymidine [( 3H]Tdr) incorporation in cultured rat FRTL-5 thyroid cells. Furthermore, to elucidate the mechanisms of human chorionic gonadotropin (hCG) in promoting the thyroid growth, we evaluated the effects of blocking type TSH receptor antibody (blocking IgGs) from patients with primary hypothyroidism on the activity of hCG. After the PEG-pretreated serum or the serum plus blocking IgGs was incubated for 72 h at 37 degrees C with FRTL-5 cells and [3H] Tdr, [3H] Tdr incorporated in the cells was counted. Although 9 normal pregnant women had normal TGA, two patients with hydatidiform mole, whose hCG levels were 966,500 and 497,100 IU/L, had positive TGA, but the activity showed normal when analyzed with the addition of a blocking IgG. hCG also showed a dose-dependent increase in [3H]Tdr incorporation, and it was inhibited by the addition of blocking IgGs. Furthermore, the inhibition of hCG-induced [3H]Tdr incorporation by 16 blocking IgGs correlated with their TBII and the inhibition activity of hCG-induced cAMP accumulation. Analysis by the Lineweaver-Burk plots of dose response curves of TSH- and hCG-induced [3H]Tdr incorporation showed the same inhibition pattern as with the addition of the same blocking IgGs. In conclusion, 1) hCG-related TGA exists in the sera of some patients with hydatidiform mole; and 2) hCG and the sera of some patients with hydatidiform mole promote thyroid growth, at least in a part, via TSH-receptors in FRTL-5 cells.     

Cytological distribution of chorionic gonadotropin subunit and placental lactogen messenger RNA in neoplasms derived from human placenta

Normal trophoblast of the human placenta elaborates at least two major protein hormones, chorionic gonadotropin (hCG), and placental lactogen (hPL). There are several gestational trophoblastic diseases of the placenta called hydatidiform mole, invasive mole, and choriocarcinoma. Molar and choriocarcinoma tissues characteristically synthesize large amounts of hCG and small quantities of hPL. To examine the role of trophoblast differentiation in the expression of the hCG and hPL genes, we studied the cytological distribution of their messenger RNA (mRNA) in tissue sections of human hydatidiform mole and choriocarcinoma by in situ hybridization. Histologically, these tissues are in different stages of cellular differentiation. In normal placenta, hCG alpha and - beta mRNA can be localized to some cytotrophoblasts and primarily to the syncytium, whereas hPL mRNA appears only in the syncytial layer. In hydatidiform mole, which still retains placental villous morphology, the hPL gene and hCG alpha and -beta genes are expressed but are poorly localized because of the admixture of cyto- and syncytiotrophoblasts. By contrast, choriocarcinoma, which is devoid of placental villous pattern but in which the cyto- and syncytiotrophoblast-like components are distinguishable, expresses hCG alpha and -beta in the syncytial- like areas but little, if any, hPL. These results suggest that a certain level of trophoblast differentiation, such as villous formation, is associated with hPL expression, while the hCG alpha gene and the hCG beta gene can be expressed in more disorganized tissues that contain cytotrophoblastic elements.     

Inhibin: a new circulating marker of hydatidiform mole?

OBJECTIVE--To define the concentrations of inhibin in serum and tissue of patients with hydatidiform mole and assess their value as a clinical marker of the condition. DESIGN--Prospective study of new patients with hydatidiform mole, comparison of paired observations, and case-control analysis. SETTING--A university hospital, two large public hospitals, and a private women''s clinic in Japan. PATIENTS--Seven consecutive referred patients seen over four months with newly diagnosed complete hydatidiform mole, including one in whom the mole was accompanied by viable twin fetuses (case excluded from statistical analysis because of unique clinical features). All patients followed up for six months after evacuation of molar tissue. END POINT--Correlation of serum inhibin concentrations with trophoblastic disease. MEASUREMENTS AND MAIN RESULTS--Serum concentrations of inhibin, human chorionic gonadotrophin, and follicle stimulating hormone were compared before and seven to 10 days after evacuation of the mole. Before evacuation the serum inhibin concentrations (median 8.3 U/ml; 95% confidence interval 2.4 to 34.5) were significantly greater than in 21 normal women at the same stage of pregnancy (2.8 U/ml; 2.1 to 3.6), and inhibin in molar tissue was also present in high concentrations (578 U/ml cytosol; 158 to 1162). Seven to 10 days after evacuation inhibin concentrations in serum samples from the same patients declined significantly to values (0.4 U/ml; 0.1 to 1.4) similar to those seen in the follicular phase of normal menstrual cycles. None of the four patients whose serum inhibin concentrations were 0.4 U/ml or less after evacuation developed persistent trophoblastic disease. Though serum human chorionic gonadotrophin concentrations declined after evacuation (6.6 x 10(3) IU/l; 0.8 x 10(3) to 32.6 x 10(3], they remained far higher than in non-pregnant women. Serum follicle stimulating hormone concentrations remained suppressed. CONCLUSIONS--In this small study serum inhibin concentrations higher than those found in the early follicular phase one to two weeks after evacuation of a hydatidiform mole seemed to be specific for persistent trophoblastic disease. Further data are needed to confirm these promising results.     

Failure of contraceptive steroids to modify human chorionic gonadotrophin secretion by hydatidiform mole tissue and choriocarcinoma cells in culture

The effect of steroids contained in oral contraceptives, namely ethinylestradiol:17α-ethinyl-1,3,5(10)-estratriene-3, 17-diol (E) and norethindrone acetate: 17β-acetoxy-17-ethinyl-4-estren-3-one (N), on cell replication and human chorionic gonadotropin (hCG) secretion by choriocarcinoma cells in monolayer culture and by hydatidiform mole tissue maintained in organ culture were studied. The steroids were added to the culture medium individually or in combination to achieve a range of concentrations (10^?10 to 10^?4), within and beyond the presumed concentration of these substances in the blood of women taking oral contraceptives. The effect of luteinizing hormone releasing hormone (LHRH) on hCG secretion by choriocarcinoma cells in monolayer culture also was investigated. The rate of hCG production by either choriocarcinoma cells in monolayer culture or by hydatidiform mole tissue maintained in organ culture was not affected by the hormones used in this study; indeed hCG secretion remained reasonably unchanged even with high concentrations of steroids (up to 10^?14 M) or LHRH (up to 10^?4 mg × ml^?1). Cell replication, as measured by increase in amount of cellular protein and DNA, was not stimulated by either of these compounds.     

P16抑癌基因在人完全性葡萄胎和正常胎盘组织中的表达

目的研究P16抑癌基因与葡萄胎发生的关系。方法分别取完全性葡萄胎和正常早孕流产标本各30例,用SABC免疫组织化学染色方法,检测P16抑癌基因在两种组织中的表达,并采用图像分析技术,对正常早孕绒毛组和葡萄胎组P16抑癌基因的表达情况进行对比分析。结果与正常绒毛相比,P16抑癌基因在完全性葡萄胎组织中的表达部位和表达量有显著性差异。结论P16抑癌基因与人完全性葡萄胎发生密切相关。     

A controlled trial of Bestatin in hydatidiform mole

Summary A prospective randomized controlled trial was conducted to study whether Bestatin, an immunomodifier, can reduce the incidence of persistent gestational trophoblastic disease in patients with hydatidiform mole. A group of 21 patients (Bestatin group) received 30 mg Bestatin daily after evacuation of the hydatidiform mole. A second group of 23 patients (control group) did not receive any drug. Blood was taken for white cell counts, differential counts, lymphocyte subset counts (CD2⁺, CD4⁺, CD8⁺ and B cells) and natural killer cell activity before evacuation of the hydatidiform moles. The tests were repeated every 4 weeks after evacuation until the serum subunit of human chorionic gonadotropin (hCG) had returned to normal or until the patient had to receive chemotherapy because of persistent gestational trophoblastic disease. There was no significant difference in the age of the patients, the pre-evacuation serum hCG, or the gestational age between the two groups. Chemotherapy was needed by 6 patients in the Bestatin group (28.6%) and 3 patients in the control group (13%) because of persistent gestational trophoblastic disease. There was no significant difference in any of the immunological parameters between the two groups before or after evacuation. We conclude that Bestatin at this dosage does not improve the immunological functions or clinical outcome in patients with hydatidiform mole.     

The clinical evaluation of the simultaneous measurements of human chorionic gonadotropin (hCG) and its alpha-subunit in sera of patients with trophoblastic diseases

The concentrations of human chorionic gonadotropin (hCG) and its free immunoreactive alpha-subunit (hCG-alpha) in the sera of patients with trophoblastic diseases were measured by hCG and hCG-alpha radioimmunoassay (RIA), respectively. In the sera of 12 women with hydatidiform mole large amounts of hCG and considerably high level of hCG-alpha were detected in all cases. After the evacuation of mole the serum level of these glycoproteins decreased, the leve of hCG-alpha declined more rapidly than hcg. in the sera of patients with destructive mole the concentration of hCG-alpha was usually lower than that of hCG. After hysterectomy and chemotherapy the levels of hCG-alpha declined practically paralleling that of hCG. However, when hCG had decreased to undetectable level, hCG-alpha could no longer be detected in all cases. Although in the serum of patient with choriocarcinoma involving the uterus and lungs the concentration of hCG-alpha was almost as high as that of hCG, the secretory pattern of hCG and hCG-alpha might not be closely related. The changes in the serum level of free hCG-alpha as well as that of hCG parelled the clinical course of the patients examined in this study. The present results suggest that measurements of the serum free hCG-alpha may be a useful parameter to follow the clinical course and to evaluate the efficacy of treatments of trophoblastic diseases.     

Comparative study of placental protein 19, human chorionic gonadotrophin and pregnancy-specific β1-glycoprotein as immunohistochemical markers for extravillous trophoblast in pregnancy and trophoblastic disease

Summary PP19, a new placental tissue protein, has 1-1 electrophoretic mobility, a molecular weight of 36 500 and 3.9% carbohydrate. To study immunocytochemical PP19 localization in extravillous trophoblast, we obtained formalin-fixed specimens from extravillous tubal pregnancy at gestational weeks (GW) 7–9 (12 blocks); four early intrauterine pregnancies at GW 7–13 (12 blocks); four late pregnancies at GW 28–38 complicated with intramural uterine myoma, placenta increta and abruptio placenta (8 blocks); four invasive complete moles (9 blocks); and seven primary and metastatic gestational choriocarcinomas (12 blocks). Immunohistochemical staining was done for PP19, pregnancy-specific 1-glycoprotein (SP1) and human chorionic gonadotrophin (hCG) using the indirect-labeled antibody method [purified PP19 (Lot no. 225/242) and antibody against PP19 (Lot no. 632ZA) prepared by H. Bohn, antibodies against hCG (Behringwerke, Marburg, FRG) and SP1 (Dakopatts, Copenhagen, Denmark)]. In both early and late intrauterine pregnancies, the extravillous syncytiotrophoblastic cell (XST) showed positive staining for hCG and SP1 in the cytoplasm, as well as for PP19, which stained more intensively in the nucleus than in the cytoplasm. The three proteins were not seen in the evtravillous cytotrophoblastic cell (XCT) in the trophoblastic cell column and shell. The interstitial cytotrophoblast-like cell (ICT), which infiltrated into the decidua and myometrium, and their blood vessels, was immunoreactively positive for PP19 but negative for hCG and SP1 with the exception of SP1-positive ICT in the myometrium in late pregnancy. XST and ICT in the endosalpinx of tubal pregnancy stained for all three proteins. In invasive complete mole, XST stained for the three proteins, but ICT infiltrating into the decidua and myometrium stained more intensively for PP19 than for either hCG or SP1. XCT did not stain for the three proteins. Staining for the three proteins in gestational choriocarcinoma resembled that in invasive mole. By PP19 staining, XST and ICT infiltrating into surrounding tissue were clearly distinguishable from other cells of similar shape. PP19 staining thus can be a useful histochemical marker in assessing the cell viability of the trophoblastic tumor after chemotherapy.     

Glycodelin protein and mRNA is downregulated in human first trimester abortion and partially upregulated in mole pregnancy.

Glycodelin (Gd) is a major reproductive glycoprotein and a mediator for immunomodulatory effects directed to cellular, humoral, and innate immunity. Human pregnancy depends on a diversity of physiological processes including modulation of the maternal immunosystem. We evaluated the expression of Gd protein and mRNA in first trimester decidual tissue of normal pregnancies and spontaneous abortion and hydatidiform moles. Furthermore, in vitro experiments on endometrial cancer cells to analyze the effect of human chorionic gonadotropin (hCG) on Gd regulation were performed. In decidual tissue of abortion patients, Gd expression was significantly decreased compared with normal gestation, which was confirmed by in situ hybridization. In mole pregnancy, an upregulation of Gd in the first 8 weeks of pregnancy was present. Gd is a main product of decidual tissue in the first trimester of human pregnancy. Reduced Gd expression in abortive pregnancy could lead to an increased activation of the maternal immunosystem, thus causing rejection of the developing fetus. Moreover, Gd expression in endometrial cancer cells in vitro could be stimulated by addition of hCG. Therefore, we speculate that hCG could be one of the factors regulating Gd expression because hCG is downregulated in women with abortion and upregulated in mole pregnancy. In addition, we found a positive feedback loop in Gd and hCG expression in human pregnancy.     

磁性分离酶联免疫技术检测人类绒毛膜促性腺激素及其临床意义分析

应用磁性分离酶联免疫检测技术（ＭＡＬＡ）对２１０名正常非妊娠妇女的血清进行ＨＣＧ定量测定,平均值３．５４９ｍＩＵ／ｍｌ血清。另外,对１０名不全流产患者、１５名葡萄胎患者及５名早孕误诊患者血清进行ＨＣＧ定量测定,均作出准确诊断。ＨＣＧ定量测定在临床诊断上有重要意义。该法具有灵敏、快速、准确、无污染等特点。是目前临床生殖内分泌激素定量测定的好方法。     

Structures, function, and transformational changes of the sugar chains of glycohormones

Human chorionic gonadotropin (hCG), human luteinizing hormone, human thyroid-stimulating hormone, and human follicle-stimulating hormone are closely related family of proteins which share a common alpha-subunit. However, their sugar moieties are quite different. hCG contains five acidic asparagine-linked sugar chains. These five sugar chains are derived by sialylation from three neutral oligosaccharides: two biantennary (N-1 and N-2) and one monoantennary (N-3) complex-type oligosaccharides. Although hCG purified from the urine of pregnant women is more enriched in sialylated sugar chains than that purified from placenta, the molar ratio of N-1, N-2, and N-3 of these two hCGs are the same (1:2:1). Comparative study of the sugar moieties of the alpha- and beta-subunits of hCG revealed that alpha contains 1 mol each of N-2 and N-3, while beta contains 1 mol each of N-1 and N-2. This specific distribution of oligosaccharides at the four asparagine loci of the hCG molecule is now helping us to consider the functional role of the sugar moiety of glycohormones. hCG is produced not only by the trophoblast but also by various trophoblastic diseases. The hCGs purified from the urine of patients with hydatidiform mole contain the same oligosaccharides as normal hCG. However, those from the urine of choriocarcinoma patients contain five additional neutral oligosaccharides. In contrast, hCGs from invasive-mole patients contain three of the five oligosaccharides, specifically found in choriocarcinoma hCGs. The malignant transformational change of the sugar moiety of hCG can be explained by an increase of a fucosyltransferase, which forms the Fuc alpha 1----6GlcNAc group and by ectopic expression and subsequent modification of N-acetylglucosaminyltransferase IV. The appearance of tumor-specific sugar chains of hCG has been used to develop a new diagnostic method for invasive mole and choriocarcinoma.     

完全性葡萄胎和正常胎盘组织中p21的表达

目的研究p21表达与葡萄胎发生的关系。方法取完全性葡萄胎和正常早孕流产标本各30例,用SABC免疫组织化学染色方法,检测p21癌基因在两种组织中的表达,并采用图像分析技术,对正常早孕绒毛组和葡萄胎组织p21癌基因的表达情况进行对比分析。结果与正常绒毛相比,p21癌基因在葡萄胎组织中的表达量没有显著性差异,表达部位有明显不同。结论p21癌基因与完全性葡萄胎的发生密切相关。     

The hydatidiform mole

ABSTRACT

The hydatidiform mole (HM) is a placental pathology of androgenetic origin. Placental villi have an abnormal hyperproliferation event and hydropic degeneration. Three situations can be envisaged at its origin: 1. The destruction/expulsion of the female pronucleus at the time of fertilization by 1 or 2 spermatozoa with the former being followed by an endoreplication of the male pronucleus leading to a complete hydatidiform mole (CHM) 2. A triploid zygote (fertilization by 2 spermatozoa) leading to a partial hydatidiform mole (PHM) but can also lead to haploid and diploid clones. The diploid clone may produce a normal fetus while the haploid clone after endoreplication generates a CHM 3. A nutritional defect during the differentiation of the oocytes or the deterioration of the limited oxygen pressure during the first trimester of gestation may lead to the formation of a HM.

In countries with poor medical health care system, moles (mainly the CHM) can become invasive or, in rare cases, lead to gestational choriocarcinomas.     

Heterogeneity of human chorionic gonadotropin in various biological fluids as revealed by chromatofocusing

This study demonstrates that chromatofocusing is powerful in analyzing multiple forms of hCG from biological fluids. For analyzing hCG from biological fluids, it is necessary to perform chromatofocusing, in the range of pH 6.2-3.0 and pH 9.0-6.0. By chromatofocusing, highly purified hCG (CR121) was found to be acidic, in the range of pI 4.22-3.8, and hCG beta was more acidic, in the range of pI 4.0-3.2. Moreover, hCG from the first trimester pregnancy, hydatidiform mole or choriocarcinoma was also mainly acidic. Therefore, chromatofocusing in the range of 6.2-3.0 was suitable for analyzing purified hCG, hCG beta, and urinary hCG from the first trimester pregnancy, hydatidiform mole and choriocarcinoma. On the other hand, because hCG in the third trimester pregnancy and the toxemia of pregnancy were mainly alkaline, the chromatofocusing system in the range of pH 9.0-6.0 was suitable for analyzing hCG from the third trimester pregnancy and the toxemia of pregnancy.     

Droplet-digital PCR assay to detect Merkel cell polyomavirus sequences in chorionic villi from spontaneous abortion affected females

Droplet-digital polymerase chain reaction (ddPCR) technique was set up to detect/quantify Merkel cell polyomavirus (MCPyV) DNA in clinical specimens, including chorionic villi and peripheral blood mononuclear cells (PBMCs) from spontaneous abortion (SA)-affected females. This ddPCR assay showed high accuracy, sensitivity, and specificity in detecting MCPyV DNA cloned in a recombinant plasmid vector, the control. ddPCR was extended to MCPyV DNA to investigate/quantify its sequences in clinical samples. Overall, 400 samples were analyzed, that is, 100 chorionic villi and 100 PBMCs, from SA females (n = 100), the cases, and 100 chorionic villi and 100 PBMCs from females who underwent voluntary pregnancy interruption (VI, n = 100), the control. MCPyV DNA was detected in 4/100 (4%) and 5/100 (5%) of SA and VI chorionic villi, respectively. The mean viral DNA load was 1.99 ( ± 0.94 standard mean deviation [SD]) copy/10⁴ cells in SA and 3.02 ( ± 1.86 [SD]) copy/10⁴ cells in VI. In PBMCs, MCPyV DNA was revealed in 9/100 (9%) and 14/100 (14%) of SA and VI, with a mean of 2.09 ( ± 1.17 [SD]) copy/10⁴ cells and 4.09 ( ± 4.26 [SD]) copy/10⁴ cells in SA and VI, respectively. MCPyV gene expression analysis by quantitative PCR for the large T antigen (LT) and viral capsid protein 1 (VP1) showed their mRNAs in 2/4 (50%) SA- and 2/5 (40%) VI-MCPyV-positive samples. MCPyV DNA was detected/quantified using the ddPCR technique, in chorionic villi and PBMCs from SA and VI. In our experimental conditions, ddPCR provided a powerful tool to detect/quantify MCPyV DNA sequences in clinical samples.