real-time polymerase chain reaction

实时聚合酶链反应
  • 文章类型: Journal Article
    虽然实时逆转录PCR(RT-PCR)是诊断严重急性呼吸道综合征冠状病毒2(SARS-CoV-2)感染的推荐实验室方法,由于高测试需求和试剂短缺,其在资源有限的环境中的使用可能难以维持。这项研究的目的是评估RealyTech™和StandardQ™在相对较低的COVID-19患病率环境中与RT-PCR相比的性能,马里。
    我们在2021年1月至4月之间在巴马科和卡蒂地区进行了一项横断面研究,以评估在马里进行的大型SARS-CoV-2流行研究中的两种快速测试。
    在测试的390个样品中,RealyTech™和StandardQ™的敏感性和特异性为57.1%(95CI:44.1-69.2),95.8%(95CI:93.1-97.5);61.9%(95CI:46.8-75.0),和94.1%(95CI:89.5-96.8)。使用RT-PCR,SARS-CoV-2的全球患病率为14.4%(56/390)。在两种快速抗原测试中,与接受治疗的阳性患者相比,在疑似患者中使用时的表现更好.此外,相当于Ct<25的较高病毒载量与较好的检出率相关.
    在等待更完整的数据时,这些初步研究表明,RealyTech™和StandardQ™不应单独用于马里的COVID-19诊断。
    UNASSIGNED: While real-time reverse transcription PCR (RT-PCR) is the recommended laboratory method to diagnose severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) infection, its use in resource limited settings can be difficult to maintain due to high testing demand and shortage of reagents. The aim of this study was to evaluate the performances of Realy Tech™ and Standard Q™ in comparison to RT-PCR in a relatively low COVID-19 prevalence setting, Mali.
    UNASSIGNED: We conducted a cross-sectional study between January and April 2021 in Bamako and Kati regions to evaluate both rapid tests during a large SARS-CoV-2 prevalence study in Mali.
    UNASSIGNED: Of the 390 samples tested, the sensitivity and specificity of Realy Tech™ and Standard Q™ were 57.1% (95%CI: 44.1-69.2), 95.8% (95%CI: 93.1-97.5); 61.9% (95%CI: 46.8-75.0), and 94.1% (95%CI: 89.5-96.8) respectively. Using RT-PCR, the global prevalence of SARS-CoV-2 was 14.4% (56/390). In both rapid antigen tests, the performance was better when used in suspected patients compared to positive patients under treatment. Moreover, higher viral loads equivalent to Ct < 25 were associated with better detection rates.
    UNASSIGNED: While waiting for more complete data, these preliminary studies suggest that Realy Tech™ and Standard Q™ should not be used alone for COVID-19 diagnosis in Mali.
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  • 文章类型: Case Reports
    由布鲁氏菌感染引起的脊柱炎是一种罕见但具有挑战性的疾病,其成功的治疗取决于及时的诊断和适当的治疗。本研究报告两例典型的胸腰布病脊柱炎病例,强调实时聚合酶链反应(实时PCR)检测和手术干预的关键作用。
    病例1涉及一名49岁的男牧羊人,有6个月的发烧史(40°C),严重的胸部和背部疼痛,2周下肢活动受限伴夜间加重。体格检查显示T9和T10棘突有压痛和搏击痛,下肢有2级肌肉力量.CT显示T9和T10椎骨骨破坏,椎间隙变窄,而MRI显示T9-T10椎骨的异常信号,椎管脓肿,和脊髓压迫。玫瑰红平板凝集试验呈阳性。病例2是一名59岁的男子,他抱怨严重的胸腰椎背痛伴发热(39.0°C),行走受限2个月。他的体重减轻了2.5公斤,并且有与绵羊密切接触的历史。玫瑰红测试呈阳性,MRI显示L1和L2椎骨的炎性改变。诊断和治疗:实时PCR证实布鲁氏菌感染。术前使用多西环素进行抗菌治疗,利福平,头孢他啶-舒巴坦至少给药2周。手术治疗包括椎间孔切开术辅助清创术,减压,内固定,全身麻醉下植骨.术后组织病理学检查,HE和革兰氏染色进一步证实了诊断。结果:在干预后的最后一次随访(4-12周),两名患者均经历了显着的疼痛缓解并恢复正常的下肢运动。
    实时PCR检测为疑似布鲁氏菌病脊柱炎病例提供了有价值的诊断见解。手术治疗有助于控制感染,脊髓减压,恢复稳定,构成必要和有效的治疗方法。在这种情况下,及时的诊断和全面的管理对于取得有利的结果至关重要。
    UNASSIGNED: Spondylitis caused by Brucella infection is a rare but challenging condition, and its successful management depends on timely diagnosis and appropriate treatment. This study reports two typical cases of thoracic and lumbar brucellosis spondylitis, highlighting the pivotal roles of real-time polymerase chain reaction (real-time PCR) detection and surgical intervention.
    UNASSIGNED: Case 1 involved a 49-year-old male shepherd who presented with a 6-month history of fever (40°C), severe chest and back pain, and 2-week limited lower limb movement with night-time exacerbation. Physical examination revealed tenderness and percussion pain over the T9 and T10 spinous processes, with grade 2 muscle strength in the lower limbs. CT showed bone destruction of the T9 and T10 vertebrae with narrowing of the intervertebral space, whereas MRI demonstrated abnormal signals in the T9-T10 vertebrae, a spinal canal abscess, and spinal cord compression. The Rose Bengal plate agglutination test was positive. Case 2 was a 59-year-old man who complained of severe thoracolumbar back pain with fever (39.0°C) and limited walking for 2 months. He had a 2.5 kg weight loss and a history of close contact with sheep. The Rose Bengal test was positive, and the MRI showed inflammatory changes in the L1 and L2 vertebrae. Diagnosis and treatment: real-time PCR confirmed Brucella infection in both cases. Preoperative antimicrobial therapy with doxycycline, rifampicin, and ceftazidime-sulbactam was administered for at least 2 weeks. Surgical management involved intervertebral foraminotomy-assisted debridement, decompression, internal fixation, and bone grafting under general anesthesia. Postoperative histopathological examination with HE and Gram staining further substantiated the diagnosis. Outcomes: both patients experienced significant pain relief and restored normal lower limb movement at the last follow-up (4-12 weeks) after the intervention.
    UNASSIGNED: Real-time PCR detection offers valuable diagnostic insights for suspected cases of brucellosis spondylitis. Surgical treatment helps in infection control, decompression of the spinal cord, and restoration of stability, constituting a necessary and effective therapeutic approach. Prompt diagnosis and comprehensive management are crucial for favorable outcomes in such cases.
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  • 文章类型: Case Reports
    目的:我们报告了在哥伦比亚边境地区发现的首例Oropouche热。
    方法:使用多重实时聚合酶链反应(PCR),2019年登革热流行期间的基因测序和临床特征,共分析了175个样本,从通报给系统流行病学监测的病例,如登革热。
    结果:根据M和S基因组片段最大似然(ML)分析,来自莱蒂西亚的Oropouche病毒(OROV)分离株属于谱系2,与在埃斯梅拉达斯获得的样本有着共同的祖先,厄瓜多尔和Turbaco,哥伦比亚。患者:居住在莱蒂西亚市边境地区的一名妇女出现以下症状:发烧,头痛,眶后疼痛和肌痛.
    结论:这种跨境监测可用于发出有关该地区虫媒病毒传播的进出的警报,在公共卫生监测系统中经常被低估。
    OBJECTIVE: We report the first case of Oropouche fever detected in the border region of Colombia.
    METHODS: Using a multiplex real-time polymerase chain reaction (PCR), genetic sequencing and clinical characteristics during the dengue epidemic in 2019, a total of 175 samples were analysed, from cases notified to the system epidemiological surveillance such as dengue.
    RESULTS: The Oropouche virus (OROV) isolate from Leticia belongs to lineage 2 according to both M and S genome segments maximum likelihood (ML) analysis, shares a common ancestor with samples obtained in Esmeraldas, Ecuador and Turbaco, Colombia. The patient: a woman resident in the border neighbourhood of the municipality of Leticia had the following symptoms: fever, headache, retro-orbital pain and myalgias.
    CONCLUSIONS: This cross-border surveillance can be useful to give an alert about the entry or exit of arboviruses circulation in the region, which are often underreported in public health surveillance systems.
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  • 文章类型: Letter
    暂无摘要。
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  • 文章类型: Case Reports
    目的:中央牙源性纤维瘤(COF)很少见,来源于牙间充质组织的良性肿瘤,可能发生在上颌骨或下颌骨。本报告描述了患有卵圆形基底细胞癌综合征(NBCCS)的患者下颌骨的原发性和复发性COF。
    方法:一名36岁的非裔美国男性出现COF,17个月后复发。从两次事件中获得组织碎片,并获得IRB批准的签署同意书。解剖收集的组织块;一部分用福尔马林固定并石蜡包埋,另一个培养用于从原发性(COdF-1)和复发性(COdF-1a)肿瘤中分离细胞群。定量实时聚合酶链反应(qRT-PCR),免疫组织化学,和DNA测序用于原发性肿瘤和细胞群体的基因和蛋白质分析。
    结果:肿瘤的组织病理学分析显示纤维结缔组织中牙源性上皮索稀疏,和qRT-PCR分析的肿瘤和细胞群体(COdF-1和COdF-1a)检测到VIM,CK14、CD34、CD99和ALPLmRNA表达。通过免疫组织化学证实蛋白质表达。由于肿瘤血管化,CD34在原代组织中的表达高于肿瘤细胞。DNA测序提示患者存在PTCH1突变。
    结论:组织病理学,mRNA和蛋白质表达表明在PTCH1基因和NBCCS突变的患者中COF的罕见发生。
    OBJECTIVE: Central odontogenic fibromas (COF) are rare, benign tumors derived from dental mesenchymal tissue that may occur in the maxilla or mandible. This report describes primary and recurrent COF in the mandible of a patient with nevoid basal cell carcinoma syndrome (NBCCS).
    METHODS: A 36-year-old African American male presented with a COF and its recurrence 17 months later. Tissue pieces were obtained from both occurrences with IRB-approved signed consent. Collected tissue pieces were dissected; one portion was formalin-fixed and paraffin-embedded, and the other was cultured for the isolation of cell populations from the primary (COdF-1) and recurrent (COdF-1a) tumors. Quantification real-time polymerase chain reaction (qRT-PCR), immunohistochemistry, and DNA sequencing were used for gene and protein analysis of the primary tumor and cell populations.
    RESULTS: Histopathologic analysis of the tumor showed sparse odontogenic epithelial cords in fibrous connective tissue, and qRT-PCR analysis of tumor and cell populations (COdF-1 and COdF-1a) detected VIM, CK14, CD34, CD99 and ALPL mRNA expression. Protein expression was confirmed by immunohistochemistry. CD34 expression in primary tissues was higher than in tumor cells due to tumor vascularization. DNA sequencing indicated the patient had PTCH1 mutations.
    CONCLUSIONS: Histopathology, mRNA, and protein expression indicate the rare occurrence of COF in a patient with mutated PTCH1 gene and NBCCS.
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  • 文章类型: Journal Article
    背景:正常的癌前倾向,胰腺未受影响的部分没有像胰腺癌病例中记录的多中心那样得到很好的探索。为了确定炎症标志物和红细胞癌基因B(ErbB2)在胰腺癌患者未受影响的胰腺中的表达,进行了一项病例对照研究.材料和方法在接受胰十二指肠切除术治疗胰腺癌(PC)的患者中,促炎基因和肿瘤标志物,使用qRT-PCR在正常胰腺切面的胰腺组织中分析了表皮生长因子受体家族中的红细胞癌基因2(ErbB2)。选择20例Frey手术后诊断为慢性胰腺炎(CP)的患者,和他们的胰腺组织作为对照进行分析。使用国家生物技术信息中心(NCBI)软件设计HPLC纯化的引物。使用基本局部比对搜索工具(BLAST)验证引物的特异性用于基因表达分析。使用β-肌动蛋白作为管家基因对研究中的基因进行归一化,并使用2-ddct方法计算与对照样品相比的倍数变化。结果未纳入切缘阳性患者。促炎基因(TNF-α,NF-kβ,与CP组相比,PC患者中COX-2)的折叠变化显着降低。CP对照组的IL-6基因表达水平高于PC组。胰腺癌患者的ErbB2基因表达明显高于CP患者。结论胰腺癌患者胰腺组织中ErbB2基因表达上调,与对照组相比,表明剩余的胰腺可能具有导致癌症的能力。原癌基因可能在胰腺癌患者的病理生理过程中起作用。
    Background The pre-malignant tendency of the normal, non-affected portion of the pancreas is not as well explored as the multicentricity documented in pancreatic cancer cases. In order to ascertain the expression of inflammatory markers and Erythroblastic Oncogene B (ErbB2) in the non-affected pancreas in patients with pancreatic cancer, a case-control study was carried out. Materials and methods In patients who underwent pancreatoduodenectomy for pancreatic cancer (PC), pro-inflammatory genes and a tumor marker, erythroblastic oncogene 2 (ErbB2) in the epidermal growth factor receptor family were analyzed in the pancreatic tissue at the cut surface of the normal pancreas using qRT-PCR. Twenty patients diagnosed with Chronic pancreatitis (CP) after Frey\'s surgical procedure were selected, and their pancreatic tissues were analyzed as controls. The HPLC-purified primers were designed using National Center for Biotechnology Information (NCBI) software. The primer\'s specificity was verified for gene expression analysis using the Basic Local Alignment Search Tool (BLAST). The genes under study were normalized using β-actin as the housekeeping gene, and the 2-ddct method was used to compute the fold change compared to the control sample. Results Patients with margin-positive were not included. Pro-inflammatory genes (TNF-α, NF-kβ, and COX-2) had significantly lower foldchange in PC patients compared to the CP group. The CP control group had higher levels of IL-6 gene expression than the PC group. Patients with pancreatic cancer had a considerably higher expression of the ErbB2 gene than patients with CP. Conclusion The upregulated ErbB2 gene in the unaffected pancreatic tissue of pancreatic cancer patients, when compared to controls, indicates that the remaining pancreas may have the capacity to cause cancer. Proto-oncogene may play a role in the pathophysiologic process in patients with pancreatic cancer.
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  • 文章类型: Journal Article
    背景:随着许多国家本土疟疾病例接近零,消除疟疾的进展正在增加。在接近消除的环境中,当针对残留的传播病灶时,干预措施将最有效地中断传播。这些病灶可能因无症状感染而错过。为了解决这个问题,世界卫生组织建议反应性病例检测(RACD).这个案例研究是为了确定无症状疟疾的个体,他们的易感危险因素,并推荐阿苏托病的RACD,加纳基于文献综述和横断面研究。
    方法:该研究涉及对PubMed和GoogleScholar在1月1日之间发表的文献进行搜索,2009-8月14日,2023年使用“Asutsuare”中的搜索词“疟疾”。此外,对100名没有疟疾症状的人进行了结构化问卷调查,并使用快速诊断测试(RDT)试剂盒进行了筛查,显微镜和实时聚合酶链反应(RT-PCR)。在横断面研究中,通过问卷调查评估了基于三种诊断技术的疟疾患病率以及潜在的疟疾危险因素。
    结果:累计,六十四(64)项研究(谷歌学者,57和PubMed,7)进行了综述,文献中包括了22项关于阿兹多拉疟疾的研究,加纳。重要的危险因素是职业,从房子到水体的距离,年龄组和教育水平。在100个样本中,3(3%)为RDT阳性,6(6%)通过显微镜和9(9%)通过rt-PCR。在4名参与者中,5-14.9岁的平均疟疾寄生虫密度最高,为560个寄生虫/μl,恶性疟原虫为优势种。此外,在≥15岁的年龄组中,2名参与者(各1名)携带恶性疟原虫和疟原虫寄生虫.RDT的灵敏度(76.54%;CI9566.82-85.54)高于rt-PCR(33.33%;CI954.33-77.72),而rt-PCR和RDT均具有较高的特异性(92.55;CI9585.26-96.95)和(97.30;CI9593.87-99.13),分别诊断为疟疾。
    结论:在白虫中,加纳,低流行区,消除疟疾可能需要寻找无症状感染的个体。鉴于本研究中确定的无症状个体的患病率较低,并且在文献综述中得到了重复,这有利于RCD,Asutsuare是RCD实施的可能设置。
    BACKGROUND: Progress toward malaria elimination is increasing as many countries near zero indigenous malaria cases. In settings nearing elimination, interventions will be most effective at interrupting transmission when targeted at the residual foci of transmission. These foci may be missed due to asymptomatic infections. To solve this problem, the World Health Organization recommends reactive case detection (RACD). This case study was conducted to identify individuals with asymptomatic malaria, their predisposing risk factors and recommend RACD in Asutsuare, Ghana based on literature review and a cross sectional study.
    METHODS: The study involved a search on PubMed and Google Scholar of literature published between 1st January, 2009-14th August, 2023 using the search terms \"malaria\" in \"Asutsuare\". Furthermore, structured questionnaires were administered to one hundred individuals without symptoms of malaria and screened using rapid diagnostic test (RDT) kits, microscopy and real-time polymerase chain reaction (rt-PCR). Malaria prevalence based on the three diagnostic techniques as well as potential malaria risk factors were assessed through questionnaires in a cross-sectional study.
    RESULTS: Cumulatively, sixty-four (64) studies (Google Scholar, 57 and PubMed, 7) were reviewed and 22 studies included in the literature on malaria in Asutsuare, Ghana. Significant risk factors were occupation, distance from a house to a waterbody, age group and educational level. Out of the 100 samples, 3 (3%) were positive by RDT, 6 (6%) by microscopy and 9 (9%) by rt-PCR. Ages 5-14.9 years had the highest mean malaria parasite densities of 560 parasites/µl with Plasmodium falciparum as the dominant species in 4 participants. Moreover, in the age group ≥ 15, 2 participants (1 each) harboured P. falciparum and Plasmodium malariae parasites. RDT had a higher sensitivity (76.54%; CI95 66.82-85.54) than rt-PCR (33.33%; CI95 4.33-77.72), while both rt-PCR and RDT were observed to have a higher specificity (92.55; CI95 85.26-96.95) and (97.30; CI95 93.87-99.13), respectively in the diagnosis of malaria.
    CONCLUSIONS: In Asutsuare, Ghana, a low endemic area, the elimination of malaria may require finding individuals with asymptomatic infections. Given the low prevalence of asymptomatic individuals identified in this study and as repleted in the literature review, which favours RACD, Asutsuare is a possible setting receptive for RACD implementation.
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  • 文章类型: Journal Article
    十二指肠贾第鞭毛虫是一种常见的致病性肠道原虫寄生虫,在发展中国家患病率较高。尤其是在儿童中。贾第鞭毛虫组合在人类中的分布及其临床相关性仍存在争议。这项研究旨在确定Jimma5岁以下儿童中贾第虫的患病率和聚集情况。埃塞俄比亚西南部。采用案例控制设计,从2016年12月至2018年7月,在Jimma大学医学中心和Serbo健康中心出现腹泻的606名儿童以及617名没有腹泻的匹配对照。使用实时PCR检测和分型贾第虫。进行单因素和多因素回归分析。贾第虫的总患病率为41%(501/1223),病例和对照组之间没有显着差异(40%vs42%)。患病率随年龄增加,在≥25个月的儿童中患病率最高。与没有腹泻史的儿童相比,上个月没有腹泻史的儿童更可能是贾第鞭毛虫阳性(OR1.8和95CI;1.1-2.9)。不管目前的腹泻症状如何,组合B占89%,其次是组合A(8%)和混合感染组合A和B(3%)。我们报告了在Jimma通过PCR检测发现贾第虫的高患病率,埃塞俄比亚,组合B占主导地位。有和没有腹泻的儿童之间贾第虫组合的分布相似。年龄增长是贾第虫感染的危险因素。需要采用基于社区的预防和控制策略来降低贾第虫感染的风险。
    Giardia duodenalis is a common pathogenic intestinal protozoan parasite with high prevalence in developing countries, especially among children. The distribution of giardia assemblages among humans and their clinical relevance remains controversial. This study aimed to determine the prevalence and assemblage of Giardia among children under 5 years of age in Jimma, Southwest Ethiopia. Employing a case-control design, 606 children presenting with diarrhea at Jimma university medical center and Serbo Health Center were enrolled from December 2016 to July 2018 along with 617 matched controls without diarrhea. Giardia was detected and typed using real-time PCR. Univariate and multivariate regression analysis was performed. The total prevalence of Giardia was 41% (501/1223) and did not differ significantly between cases and controls (40% vs 42%). Prevalence increased by age, with the highest prevalence seen in children aged ≥ 25 months. Children without diarrhea with a history of diarrhea during the last month were more likely to be Giardia positive compared to children with no history diarrhea (OR 1.8 and 95%CI; 1.1-2.9). Regardless of current diarrhea symptoms, assemblage B predominated with 89%, followed by assemblage A (8%) and mixed infection assemblage A and B (3%). We report a high prevalence of Giardia by PCR detection in Jimma, Ethiopia, with assemblage B being predominant. There was a similar distribution of Giardia assemblages between children with and without diarrhea. Increasing age was a risk factor for Giardia infection. Community-based prevention and control strategies need to be employed to decrease the risk of giardia infection.
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  • 文章类型: English Abstract
    Paracoccidioidomycosis is a systemic mycosis endemic in Latin America. The most frequent form involves a chronic compromise of the lungs, skin, and mucosa. The patient started with a single oral lesion that lasted for several years. The absence of other symptoms pointed out a possible malignant neoplasm, specifically a squamous cell carcinoma. Differentiation between both diagnoses –fungal infection and carcinoma– depends on the results of the direct examination, the histopathological study, and the initial and subsequent cultures. However, in this case, those findings were not conclusive. The coexistence of both diagnoses is frequent and increases the diagnostic challenge. After several consultations and tests, direct examination, immunodiffusion and real-time PCR findings the multifocal chronic paracoccidioidomycosis diagnosis was confirmed. This case warns about a systematical absence of clinical suspicion of endemic mycoses before the appereance of mucocutaneous lesions, which can be produced by fungi like Paracoccidioides spp, and the importance of considering those mycoses among the differential diagnoses.
    La paracoccidioidomicosis es una micosis sistémica endémica en Latinoamérica. La presentación más frecuente compromete crónicamente los pulmones, la piel y las mucosas. Al inicio, este paciente presentó, por varios años, una lesión única en la mucosa oral que, en ausencia de otros síntomas, se relacionó con una neoplasia maligna, específicamente con un carcinoma escamocelular. La diferenciación entre los dos diagnósticos se hace mediante un examen directo, un estudio histopatológico y cultivos iniciales y subsecuentes. Sin embargo, tales estudios no fueron concluyentes. Después de varias consultas y pruebas, con los resultados del examen directo, la inmunodifusión y la PCR en tiempo real se confirmó el diagnóstico de paracoccidioidomicosis crónica multifocal. Este caso alerta sobre la ausencia de sospecha clínica de micosis endémicas, dada la presencia de lesiones mucocutáneas que pueden ser producidas por hongos como Paracoccidioides spp, y la importancia de considerarlas entre los diagnósticos diferenciales.
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  • 文章类型: Case Reports
    本报告介绍了解剖学组织病理学,对自然感染利什曼原虫(Leishmania)婴儿的无症状雌性狗及其四只幼犬进行免疫组织化学和分子分析。除了常规的血清学检测,通过骨髓样本的实时PCR证实了母亲的阳性;胎盘和卵巢,然而,相同的分析结果为阴性。幼犬在使用的任何技术中都没有阳性。提示妊娠期和与女性免疫状况相关的因素导致垂直感染发生的更大变异性。
    This report presents anatomo-histopathological, immunohistochemical and molecular analyses of an asymptomatic female dog naturally infected with Leishmania (Leishmania) infantum and her four pups. In addition to routine serological tests, the mother\'s positivity was confirmed by real-time PCR of a bone marrow sample; the placenta and ovaries, however, were negative with the same analysis. The pups presented no positivity in any of the techniques used. It is suggested that the gestational period and factors related to the female\'s immunological condition cause greater variability in the occurrence of vertical infection.
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