Urine PCR Research Articles

Fasting outcomes in people with diabetes and chronic kidney disease in East London during Ramadan 2018: The East London diabetes in Ramadan survey

ObjectivesFasting in summer months for Muslim people with diabetes during Ramadan is challenging, particularly in temperate climates, where duration of fasting is prolonged. Risk of fasting may be greater in people with diabetes and chronic kidney disease (CKD). We aimed to prospectively monitor the outcomes of patients with diabetes and CKD stage 3 during Ramadan 2018 in East London. MethodsPatients with type 2 diabetes (T2D) and CKD stage 3 attending community diabetes clinics were approached prior to Ramadan 2018 to discuss fasting. Patients were risk categorised according to Diabetes and Ramadan Alliance guidelines. If they chose to fast, Ramadan education was given, and biomedical assessments were undertaken within one week prior to and one week after fasting. Outcomes between patients fasting and non-fasting groups were compared. ResultsFasting (n = 68) and non-fasting groups (n = 71) were similar apart from slightly higher insulin use in the non-fasting group. Median days fasted was 21 (range 12–29). There were no significant changes in weight, blood pressure, creatinine, glycated haemoglobin, cholesterol and urinary PCR pre- and post-Ramadan, and no significant differences between the fasting and non-fasting groups. There was no difference in adverse events (acute kidney injury, hypoglycaemia or cardiovascular events) between the fasting and non-fasting groups. ConclusionsNo significant differences were seen in clinical or biochemical parameters, or adverse events between fasting and non-fasting patients. Patients with T2D and stable CKD stage 3 may be able to fast safely during Ramadan.

Progressive multifocal leukoencephalopathy in Zambia is caused by JC virus with prototype regulatory region

There are only few documented cases of progressive multifocal leukoencephalopathy (PML) in Africa. Whether this is caused by a lack of JC virus (JCV) spread or alteration in the JCV genome is unknown. We characterized the clinical presentation, laboratory findings, and JCV regulatory region (RR) pattern of the first documented PML cases in Zambia as well as JCV seroprevalence among HIV+ and HIV- Zambians. We identified PML patients with positive JCV DNA PCR in their cerebrospinal fluid (CSF) among subjects enrolled in an ongoing tuberculous meningitis study from 2014 to 2016 in Lusaka. JCV regulatory region was further characterized by duplex PCR in patients' urine and CSF. Of 440 HIV+ patients, 14 (3%) had detectable JCV DNA in their CSF (age 18-50; CD4+ T cells counts 15-155 × 106/μl) vs 0/60 HIV- patients. The main clinical manifestations included altered mental status and impaired consciousness consistent with advanced PML. While prototype JCV was identified by duplex PCR assay in the CSF samples of all 14 PML patients, only archetype JCV was detected in their urine. All PML Zambian patients tested were seropositive for JCV compared to 46% in a control group of HIV+ and HIV- Zambian patients without PML. PML occurs among HIV-infected individuals in Zambia and is caused by CNS infection with prototype JCV, while archetype JCV strains are present in their urine. JCV seroprevalence is comparable in Zambia and the USA, and PML should be included in the differential diagnosis of immunosuppressed individuals presenting with neurological dysfunction in Zambia.

Low serum C3 level, high neutrophil-lymphocyte-ratio, and high platelet-lymphocyte-ratio all predicted poor long-term renal survivals in biopsy-confirmed idiopathic membranous nephropathy

Idiopathic membranous nephropathy (iMN) is the major cause of end-stage renal disease (ESRD). Recent guidelines suggest limiting immunosuppressants only to high risk patients for ESRD. The present study is aimed at identifying new predictors for the renal outcome of iMN patients. We conducted a retrospective cohort study covering a period from January 2003 to December 2013. We enrolled participants who had received their first renal biopsy at our medical center in Taiwan with the diagnosis of iMN. Clinical, pathological and laboratory data were collected from medical records. Analyses with Mann–Whitney U test was used for continuous variables and Chi-square test for categorical variables. The Kaplan-Meier curve was used for the analyses of patient survival and renal survival. Youden index was used for evaluating the performance of a dichotomous diagnostic test for renal and patient outcomes. Cox proportional hazard regression was used to determine factors affecting renal survival.A total of 99 patients with renal biopsy-confirmed idiopathic iMNs were enrolled. C3 level ≤114 mg/dl predicted patient outcome (p < 0.001) with good predictive power (AUC = 0.736). The univariate analysis showed that risk factors for poor renal outcome were older age (HR = 1.04, p = 0.002), high BUN (HR = 1.03, p < 0.001), poor baseline renal function (HR = 1.30 and p < 0.001 for higher serum creatinine; HR = 0.97 and p < 0.001 for higher eGFR; HR = 1.06 and p < 0.001 for urine PCR), C3 ≤ 93.4 mg/dl (HR = 2.15, p = 0.017), NLR > 3.34 (HR = 3.30, p < 0.001) and PLR > 14.48 (HR = 2.54, p = 0.003). Stage of iMN did not fully account for the risk of ESRD. This is the first evidence that serum levels of C3 ≤ 93.4 mg/dl predicted poor renal outcomes with good predictive power. Easily obtained markers, NLR > 3.34 also predicted poor renal outcomes.

Epidemiological investigation of Leptospira spp. in a dairy farming enterprise after the occurrence of three human leptospirosis cases.

An epidemiological investigation was conducted in an unvaccinated dairy farming enterprise in which three workers on one of the milking herds (Herd 1) were diagnosed with leptospirosis due to serovars Hardjo (H) (n=2) and Pomona (P) (n=1) between January and March 2015. Blood and urine samples were collected from milking cows in Herd 1 (N=230) and Herd 2 (N=400), rising one- (R1, N=125) and rising two-year-old (R2, N=130) replacement heifers, and four pigs associated with Herd 1, in March 2015. Sera were tested using the MAT for serovars H, P, Copenhageni (C), Ballum (B) and Tarassovi (T), and urine samples were tested by qPCR. Seventy-five per cent of 109 cows in Herd 1 and 36% of 121 in Herd 2 were seropositive (≥48), predominantly to H and P, and 23% of 74 cows in Herd 1 and 1% of 90 cows in Herd 2 were qPCR positive. Fifty-five per cent of 42 R2 heifers were seropositive to T. No R1 and 17% of 42 R2 heifers were qPCR positive. Subsequently, all cattle were vaccinated for H and P, and Herds 1 and 2 were given amoxicillin. After the booster vaccination, 7% of 91 in Herd 1, 2% of 82 in Herd 2 and 11% of 38 R1 heifers (sampled as R2) were PCR positive. After the amoxicillin treatment, no cows in Herd 1 and 5% of 62 cows in Herd 2 were urine PCR positive. Calves and pigs were seropositive to H, P, C and B. Vaccination and antibiotic treatment appeared effective in reducing the risk of exposure of workers to vaccine serovars. However, evidence of non-vaccine serovars indicated that workers likely remain at risk of exposure to Leptospira.

Challenges on the Follow-Up Experimental Leptospiral Infection in Sheep

Background: Leptospirosis is currently a source of significant economic losses in the agribusiness; as such, experimental studies on this infection are required to develop a better understanding of the pathogenesis, treatment, and immunoprophylaxis of the disease. Sheep may represent a good model for ruminants in such models. Despite the extent of the studies that has been conducted thus far, researchers have yet to reach a consensus on the experimental practices to apply for leptospirosis in this animal species, and several gaps in understanding remain. To bridge these gaps, the present study aimed to assess the usage of several tools for the monitoring of experimental leptospirosis in sheep.Material, Methods &amp; Results: Twelve Santa Ines sheep of different ages were each allocated to one of four groups (A, B, C, and D). The subjects in groups A, B, and C received different doses of Leptospira interrogans serogroup Icterohemorrhagiae by intraperitoneal route, 1x102, 1x105, and 1x108 respectively. Group D was the control. Hematological, biochemical and clinical parameters were evaluated daily. Serology by microscopic agglutination test (MAT) and PCR were performed to evaluate the infection status. The most remarkable clinical signs were fever (41ºC) and dehydration, and acute pain (cub). Two animals from Group C presented leukocytosis. Only those in Group C exhibited positive results according to serology, while positivity in PCR was observed in animals in groups A and C. The results of the experiment indicated that sheep may be experimentally infected and can, therefore, be used as a model for leptospirosis in ruminants. Clinical signs cannot be considered to represent a reliable parameter for evaluating the development of leptospirosis in experimentally infected sheep. We recommend the use of urine PCR and serology to confirm the infection in experimentally infected animals and daily complete blood count (CBC) as a follow-up tool.Discussion: It was observed that the clinical signs cannot be considered as a reliable parameter to evaluate the pathogenesis in experimentally infected ewes, being important to emphasize that the age of the animals does not seem to alter their susceptibility to the infection. This finding is in agreement with other experimental studies, which report that leptospirosis infection in ruminants occurs asymptomatic and subclinical. Hematological and biochemical tests proved to be adequate tools to monitor the experimental infection. Studies have shown that the complete blood count has been used to monitor the acute phase of leptospirosis and is effective in detecting anemia and leukocytosis with neutrophilia in ruminants. Despite the lack of clinical signs, the serological and molecular results confirmed the experimental infection. PCR has been used as an important tool in the diagnosis of leptospirosis. In addition, the current study is the first of its kind to use PCR to detect the carrier status in experimentally infected ewes. Despite this limitation, PCR was very effective in confirming the infection and should be considered for use in experimental studies. Sheep have been used as a good experimental model in several studies, sheep are relatively small compared to other ruminants and can be easily allocated in smaller pens and pens, facilitating the management of research and minimizing the costs of experimentation. In this context, we suggest that sheep represent a good model for the study of leptospirosis in ruminants and therefore a reliable protocol for experimental infection by leptospirosis is necessary.

Infection à cytomégalovirus chez la femme enceinte

With a worldwide prevalence of between 0.3 and 1 %, congenital cytomegalovirus (CMV) infection is the main cause of congenital infection. Long term neurological or audiological sequelae are seen in 20 to 25 % of infected new-borns. Congenital CMV may follow maternal primary infection or maternal non-primary infection, with comparable morbidity. The diagnosis of maternal primary infection can be done at 12 to 13 weeks gestation by serology using a 2 steps algorithm. IgG and IgM are first tested and in cases with positive IgG and IgM, IgG avidity is tested. There are no tools validated to achieve the diagnosis of maternal non-primary infection. It is therefore not advisable to prescribe a CMV serology to a already known as seropositive. Diagnosis of fetal infection relies on CMV PCR in amniotic fluid. The prognosis of an infected fetus is based upon imaging and fetal laboratory markers. At birth, diagnosis relies on CMV PCR in saliva or urine collected in the first 3 weeks. False positive particularly in saliva sample are possible, a confirmation in a second sample is mandatory. Retrospective diagnosis on dried blot spots of Guthrie cards is possible with varied sensitivity depending on the assays used. No antenatal treatment is completely validated. Valganciclovir treatment in neonates with CNS involvement has a proved albeit modest efficacy with improvement or stabilization of hearing.

Successful Treatment of Toxoplasma Induced Hemophagocytic Lymphohistiocytosis (HLH) after Undergoing Allogenic Stem Cell Transplantation (HSCT)

Introduction Toxoplasmosis is a well-known intracellular protozoan parasite with seroprevalence varying geographically. Relatively asymptomatic in healthy individuals, it's reactivation can be a devastating opportunistic infection in HSCT patients with rare documentation of associated HLH. Case A 25-year-old woman with Hodgkin lymphoma, refractory to multiple chemotherapies, autologous transplant and Nivolumab, who underwent haploidentical HSCT, was admitted on day +33 with persistent high-grade fevers. Blood counts were unremarkable, except for eosinophilia. She received broad spectrum antibiotics. PET/CT on day +36 demonstrated resolution of metabolic activity in multiple lymph nodes, but revealed ‘revved-up' bone marrow(Image 1). CT-chest (day +41) demonstrated widespread centrilobular nodules. Blood, urine, enteric cultures, HIV, Histoplasma studies, and respiratory viral panel PCR were negative. Pre-transplant serologies noted a positive toxoplasma IgG. Toxoplasma DNA was detected in blood on day +41; she received clindamycin, pyrimethamine, and atovaquone due to sulfa allergy but successfully underwent rapid sulfa desensitization. She developed progressive pancytopenia and hypoxemia (day +43) requiring intubation in the ICU. Ferritin was >50,000 ng/mL, and triglycerides 931 mg/dL. Bone marrow biopsy showed activated macrophages and hemophagocytosis (Image 2). She met 6/8 criteria described in HLH-2004 trial. She received dexamethasone and tocilizumab for HLH, leading to prompt reversal of HLH physiology within 72 hours of HLH-directed therapy initiation. She was extubated on day +48. She was discharged on sulfadiazine and pyrimethamine (day +60) with almost normal ferritin levels and transfusion-independency. Discussion HLH is characterized by cytopenias, extreme systemic inflammatory response, and high mortality. HLH can be triggered by infections, such as toxoplasmosis. Disseminated toxoplasmosis has a high mortality itself (>60 % treated, and 99% untreated), and HSCT recipients appear to have the highest risk. Early detection and treatment of the underlying infection is essential. Novel drugs, such as Tocilizumab, should be prospectively studied in HLH. In conclusion, HLH after HSCT should be suspected in cases of unexplained inflammatory response and cytopenias after successful engraftment; secondary causes must be explored and promptly treated.

Research Progress on BK Virus Infection after Hematopoietic Stem Cell Transplantation--Review

BK virus infection is one of the common complications after hematopoietic stem cell transplantation(HSCT), which is also one of the reasons of the hemorrhagic cystitis．In recent years, although there are more studies of the risk factors related with human BK virus infection after hematopoietic stem cell transplantation, the risk factors related with BKV-associated hemorrhagio cystitis(BKV-HC) remain to be elucidated. Diagnosis of BK virus infection is mainly based on quantitative PCR of blood or urine. An effective strategy for treatment of these patients is the adoptive transfer of T lymphocytes specific to virus-associated antigens. In this review, the progressis of diagnosis and treatment of BK virus infection after hematopoietic stem cell transplantation are briefly summarized.

Asymptomatic leptospiral infection is associated with canine chronic kidney disease.

Canine leptospirosis is characterized by an acute or chronic disease. Some dogs may act as asymptomatic carriers, keeping the agent in the renal tubules and eliminating it in the urine for an extended period. Chronic kidney disease (CKD) is multifactorial and pathophysiology has been widely discussed. The aim of the study was to investigate whether the occurrence of CKD may possibly be associated with asymptomatic leptospiral infection in dogs in endemic regions. Serology and urine PCR were performed in 16 dogs with CKD and 48 healthy dogs from an endemic area. Dogs with CKD were more frequently shedders (75%) than non-CKD animals (20.8%). Therefore, our results demonstrate that asymptomatic leptospiral infection is associated with canine chronic kidney disease and that differential diagnosis is important for dogs from endemic areas presenting CKD. The early detection of shedders, besides the obvious impact on Public health may also help to improve the animal health and avoid the development of CKD.

Seroprevalence, frequency of leptospiuria, and associated risk factors in horses in Kansas, Missouri, and Nebraska from 2016-2017.

Leptospirosis is a worldwide veterinary and public health concern, and well recognized infectious disease of horses. Seroprevalence rates vary with geography, but many studies have confirmed a high exposure rate. The correlation between seropositivity and shedding status has not been made in horses, however. The aims of this study were to use semi-nested PCR on urine from apparently healthy horses to determine period prevalence of leptospiral shedding and to correlate these findings with MAT results to establish associations with client based survey data regarding horse management and environment. Serum and free-catch urine were collected from 204 healthy horses between May 2016-December 2017. Serum was used to determine GGT, creatinine concentrations, and six serovar MAT. Urine samples were submitted for PCR testing of leptospiral 23S rRNA. Client consent and survey data were collected for all subjects. Potential risk factors included drinking water source, exposure to livestock and dogs, geographical location, season, and precipitation. Two horses were positive on urine PCR for leptospirosis (shedding prevalence 1%), yet only one had a high reciprocal MAT titer of ≥ 800. Both horses were negative on urine PCR one month later without treatment. Approximately 77% of horses (157/204) were seroreactive (MAT reciprocal titer ≥ 100) with titers to serogroup Australis detected more frequently than others (47.5%; (97/204)). Apparently healthy horses infrequently shed Leptospira spp. in urine, yet seroreactivity in clinically normal horses is high (77%), confirming high exposure rates to Leptospira spp. in the Central Midwest.

382. Novel Influenza A/H1N1/2009 viral infection in pregnancy: A case report from Singapore

Seasonal Influenza is common in Singapore, a tropical country, with two peaks a year(April–June and year end). The novel influenza A virus is easily transmissible via respiratory droplets. High-risk population include the elderly, children, and pregnant mothers. The patient was a 40-year-old Malay lady, G5P4, a chronic hypertensive not compliant with medications. She presented at 37-weeks gestation to the emergency room with a 2-day history of hyperpyrexia, breathlessness, flu-like symptoms, and decreased fetal movement. Her vital signs were pulse rate 110, respiratory rate 20, blood pressure 190/110, temperature 39.8 °C and oxygen saturation 94–96%. On examination, her throat was injected and she had bilateral lung crepitations. Her white cell count was 14.67, CRP 64. Chest X-ray revealed patchy alveolar shadowing in the right lower zone suggestive of pneumonia. Uric acid was 594, aspartate transaminase was 84 and urine PCR was 0.3. CTG showed fetal tachycardia of 170 with poor variability. She was admitted to the high dependency labour ward for severe pre-eclampsia and possible chest infection. High flow oxygen was given together with Ceftriaxone, Azithromycin for possible chest infection and Labetalol for blood pressure control. Respiratory throat swabs were sent for Influenza A PCR and tested positive for Novel Influenza A/H1N1/2009. She was referred to Infectious Disease physicians and started on Oseltamivir. IV Magnesium Sulphate was started for her severe pre-eclampsia and she underwent induction of labour. She delivered vaginally and the newborn was admitted to high dependency neonatal ward. The patient was admitted for a total of 6 days. Pregnant women, especially those with comorbidities, are at higher risk of influenza infections, resulting in increased morbidity and mortality to mother and her newborn. Despite convincing evidence of both safety and benefit of influenza vaccination, vaccination rates among pregnant women remain low. Education of pregnant mothers and healthcare workers are essential.

Screening for Leptospira DNA in blood and urine from 30 apparently healthy dogs

Summary Asymptomatic carriage of leptospires in dogs has not been extensively studied. A prospective study was conducted on thirty apparently healthy male dogs to screen for Leptospiral DNA in their blood and urine. All dogs were up to date with a bivalent vaccine against Leptospirosis. Microscopic agglutination test (MAT) and real-time PCR analysis in blood and urine were performed for each dog. Six of 30 (20%) dogs had positive PCR results: two from blood samples (all MAT titres ≤ 160) and four from urine samples (one with maximal MAT titres of 2560 and three with all MAT titres ≤ 160). Ten months later, PCR analyses of blood and urine were repeated for these six dogs. Four had negative PCR results from blood and urine. The other two, with positive urine results at the first sampling, had negative PCR results from urine and positive PCR results from blood. In this study, the urinary shedding of leptospires appeared to be transient. As shown previously, serological status was not correlated with PCR results. Low but detectable amounts of Leptospiral DNA in the blood of some apparently healthy dogs raises questions regarding the pathophysiology of Leptospirosis and the diagnostic consequences of high-sensitivity PCR assays.

Post-Zika Guillain-Barre Syndrom: Description of clinical, biological, functional and electrophysiological characteristics observed during the Zika epidemic in Martinique

The objective is to carry out a descriptive study listing the clinical, biological, functional and electrophysiological characteristics observed in patients with post-ZIKA Guillain-Barre syndromes (SGB) during the ZIKA epidemic in Martinique, and hospitalized in a rehabilitation service. A retrospective study of subjects with post-ZIKA SGB from December 2015 to September 2016 in Martinique diagnosed according to Brighton criteria associated with molecular biology techniques, serology, and electrophysiology. Collection of data at nadir and discharge from hospitalization. Twenty-five subjects were included with a sex ratio of 1.27 and a median age of 65 years. In total, 13 patients were hospitalized in intensive care unit. At nadir, 21 patients (84%) were tetraparetic, 10 (40%) had facial diplegia, 15 (60%) had severe swallowing disorders, 12 (48%) were dysuric, 6 (24%) had acute urinary retention. Twenty-four patients (96%) had paresthesias, 11 (44%) had severe constipation requiring repeated enemas, 8 (32%) had infectious pneumonia with 7 (28%) pneumopathies with mechanical ventilation, 2 (8%) had a cardiorespiratory arrest. The median for the duration of the extension phase was 8 days versus 4 days for the plateau phase. The median for the delay between the first signs of ZIKA infection and the first neurological signs was 2.5 days. Sixteen urine PCR returned positive for ZIKA and serum neutralization was required for 9 subjects, Twenty-one acute inflammatory demyelinating polyradiculoneuritis were objectified. In total, 13 patients (52%) required a nasogastric tube, 4 (16%) required percutaneous gastrostomy, 9 (36%) had orotracheal intubation, and 6 (24%) had tracheotomy. All the patients returned home with a quasi-complete autonomy. The large number of disorders of swallowing and dysautonomia haemodynamic and vesicospincterian will be important elements to be favored in the acute management in the event of new epidemics due to their therapeutic urgency and residual incapacities. Despite severe clinical forms, patients presented good functional recovery.

Prevalence of Chlamydia Trachomatis and Neisseria Gonorrhea among Pregnant Adolescents Screened Routinely in the Third Trimester Using a Urine Pcr Test for Diagnosis: a Retrospective Chart Review

Objectives: Current Canadian antenatal care guidelines include screening for chlamydia and gonorrhea (C&G) in the first trimester for all women and repeat third trimester screening for adolescents (<21 years). This study aims to assess the rate of new and recurrent C&G infections in a multidisciplinary Adolescent OB Clinic. We hypothesize it would be appropriate to do targeted repeat screening in the adolescent population.

Arsenicosis in bladder pathology and schistosomiasis in Eggua, Nigeria.

Chronic schistosomiasis and arsenic exposure through drinking water are some of the risk factors for bladder cancer. To determine the association of schistosomiasis and arsenicosis with bladder pathologies, 122 individuals from Eggua in southwest Nigeria were recruited for this study. Prevalence of schistosomiasis was determined by urine microscopy and PCR. Total urinary arsenic concentration and arsenic levels in three different water sources in the community were assessed by flame atomic absorption spectrometry. Bladder pathologies were investigated by ultrasonography. The data collected were evaluated with chi-square (χ2) and ANOVA tests to examine the relationships among demographic factors, infection, bladder pathologies and urinary arsenic concentrations. The prevalence and mean intensity of schistosomiasis were 21.3% and 20.7 eggs/10 mL urine, respectively. Arsenic concentration in two of the water sources, River Yewa (0.46 mg/L) and borehole (0.52 mg/L), were above the WHO standard (0.01 mg/L); and the mean concentration in urine samples, 1.17 mg/L, was also above the WHO standard (0.2 mg/L). There was no evidence of an association between bladder pathology and arsenicosis, or between schistosomiasis associated-bladder pathology and arsenicosis (p=0.66). Arsenicosis is a public health concern in the study population. At the moment no clear roles are envisaged for it in the development of bladder pathologies or urinary schistosomiasis-associated bladder pathologies in Eggua.

Comparison of Culture and PCR Methods for Diagnosis of Group B &lt;i&gt;Streptococcus&lt;/i&gt; in Women

Group B Streptococcus (GBS), as a risk factor for newborns, is one of the most important causes of meningitis and septicaemia in newborns, intrauterine infections in women and colonization in the vaginal region in late gestation. We evaluate GBS prevalence and infection among pregnant women through culture and PCR methods, and then compare these methods. To this end, vaginal and urine swabs were separately collected from 246 women at the Women’s Hospital of Sari, Iran. Next, the samples were enriched in selective culture media Todd–Hewitt broth for 24 h at 37°C, recognized using blood agar media, and finally were amplified STREP gene by PCR technique. The indicated that the frequency of GBS in samples collected from urine and vaginal cultures and PCR method was positive. In addition, no significant relationship was found among the positive results of culture, maternal age, gestational age, a history of abortion and infection. With regard to the cultivation method as a standard technique, the sensitivity of PCR test was 100% and specificity was 96%. Moreover, it was found that the colonization rate of GBS in women was significant in Sari. Therefore, PCR is recommended as a reliable and rapid method for detection of GBS.

Neonates with congenital Cytomegalovirus and hearing loss identified via the universal newborn hearing screening program

BackgroundCongenital cytomegalovirus (CMV) is the most common non-genetic cause of sensorineural hearing loss. Currently, there are no universal CMV screening programs for newborns or routine CMV testing of neonates with hearing loss in Australia, or elsewhere. ObjectivesThis study was undertaken to determine the prevalence of congenital CMV infection in infants with hearing loss identified using routine resources via the Australian universal neonatal hearing screening (UNHS) program. Study designInfants who failed UNHS, referred for audiological testing and found to have permanent hearing loss were screened for CMV via PCR of urine and saliva. Congenital CMV was diagnosed if CMV was detected in infants ≤30 days of age, or using retrospective testing on stored new born screening cards, retrospective testing, or using clinical criteria if >30 days of age. The cohort was analyzed for time of testing and prevalence of congenital CMV determined. ResultsThe Audiology Department reviewed 1669 infants who failed UNHS between 2009 and 2016. Thirty percent (502/1669) had permanent hearing loss confirmed, of whom 336/502 were offered CMV testing. A definite (n = 11) or probable (n = 8) diagnosis of congenital CMV occurred in 19/323 (5.9%), of whom definite diagnoses were made in 4/19 on tests positive prior to 21 days of life, in 5/19 who were positive on neonatal blood screening card (NBSC) testing, in 2/19 who were positive on placental testing. In 8/19 probable diagnoses were made based on positive testing between ages 23–42 days and a consistent clinical syndrome in the absence of another cause for hearing loss after genetic and other testing. CMV testing mirrored the timing of audiological testing, with ∼40% completing audiology and CMV testing by 21 days, and 64% by 30 days. ConclusionThis program, utilizing existing clinical services identified probable congenital CMV in ∼6% of a large cohort failing UNHS with permanent hearing loss, of whom more than half were definite diagnoses. No additional assets were required to those already existing in this tertiary referral pediatric centre, whilst providing useful and timely data for clinical and audiological management.

A comparison of spot urine protein-creatinine ratio with 24 hour urine proteinexcretion for prediction of proteinuria in preeclampsia.

Objective: To examine the correlation between spot urine PCR and 24-hour urine protein excretion in patients being evaluated for preeclampsia. Population or sample: 98 pregnant women with suspected preeclampsia and a gestational age more than 20 weeks. Methods: A random urine sample was collected immediately before or after the 24-hour urine collection. The protein creatinine ratio was calculated by dividing the urinary protein concentration by the urinary creatinine concentration, and the correlation with the 24-hour urine protein excretion was assessed. Results: PCR at a cut-off value of 0.02 g/mmol (0.18 g/g) had sensitivity and a specificity of 97.6% and 44% respectively in predicting proteinuria of 300 mg/24 hour. The positive predictive value was 58%, and negative predictive value was 96%. Conclusion: There is strong correlation between spot PCR and 24-hour urine protein collection. PCR could be a reasonable alternative to clinicians in order to reduce their dependence on the 24-hour urine collection.

Analytical and Clinical Performance of a Real-time Screening PCR Assay Identifying Congenital CMV Infection

BackgroundCongenital CMV infection (cCMV) is the most common identifiable cause of mental retardation in the United States but requires early diagnosis to define the infection and to institute effective antiviral therapy. Traditional identification strategies including hearing screens and physical exams likely miss many patients with cCMV. We therefore developed and evaluated the performance of a PCR assay optimized for low cost, specimen collection at time of dried blood spot collection, and detection thresholds below the salivary CMV concentrations known to occur in cCMV patients.MethodsWe utilized a real-time CMV PCR assay (SimplexaTM CMV)(DiaSorin, Cypress CA) amplifying the UL83 gene and the 3M Integrated Cycler. Saliva was collected from volunteers (Copan swab), and spiked with known concentrations of CMV culture supernatant quantified by COBAS AmpliprepTM. (Roche Diagnostics). Additionally, saliva was collected by copan swab from all births within a single multi-hospital system from 3/21/16 – 5/4/17. Newborns who were initial screen PCR positive were subsequently evaluated by urine CMV PCR by an outside laboratory for confirmation of cCMV.ResultsAnalytical threshold of detection was well below 4 log copies/ML, with 100% of samples testing positive at 3.5 log copies/ML (Fig 1). 6127 newborn saliva samples were evaluated and 61 were PCR positive (£40 CT). 47 of these tests were confirmed by urine PCR (Fig 2) (PPV 0.9792, NPV 0.9988, Sens 0.8704, Spec 0.9998). Screen positive tests which were not confirmed by urine PCR had CT values £36. Adjusting the definition of a positive to CT £36 further improved the performance (PPV >0.9999, NPV 0.9997, Sens 0.9592, Spec >0.9999).ConclusionWe demonstrate good performance of a congenital CMV methodology thus facilitating an effective universal newborn screening programDisclosures J. P. Devincenzo, AstraZeneca/MedImmune: Investigator, Research support

Outcomes of women with laboratory evidence of Zika infection in pregnancy

BackgroundZika virus (ZIKV) infection in pregnancy is a global health concern. With onset of local transmission, obstetricians in Miami-Dade County, FL, United States, are now in the unique position of providing care to both pregnant women with locally-transmitted and travel-associated ZIKV infections. This study provides data regarding the testing and pregnancy outcomes of women with laboratory evidence of ZIKV infection in pregnancy.MethodsA retrospective chart review was conducted using laboratory records of ZIKV testing (PCR and IgM) completed from January through December 2016 at multiple tertiary care centers located in Miami-Dade County. Testing was based on CDC guidelines at time of testing, leading to heterogeneity in tests performed. Data was extracted from charts of women with positive ZIKV PCR in serum and/or urine or positive ZIKV IgM with confirmatory, pending, or insufficient PRNT results. Routine obstetrics parameters and the presence of fetal or neonatal abnormalities were recorded.ResultsOf the 2327 pregnant women screened for ZIKV, 88 (3.8%) screened positive with PCR and/or IgM in serum or urine. Of those women with positive ZIKV testing, 53 (60%) had no documented ZIKV symptoms and 40 (45%) had no known travel history outside of Miami-Dade County during their pregnancy. Sixty-six women had antenatal ultrasounds, 14 (21%) of which ever had a head circumference or biparetial diameter measurement less than the third percentile, but none showed evidence of intracranial calcifications. Fifty-four women with positive testing have delivered: 46 at term and 8 preterm. Fifty-four infants have been born to women with positive ZIKV testing; 2 infants (1.98%) had documented congenital abnormalities. One infant was born with clinically-defined microcephaly (1.9%) and intracranial calcifications and the other had only intracranial calcifications. Ninety-four positive IgM tests were sent to the CDC for confirmatory plaque reduction neutralization testing (PRNT). 49 PRNT tests returned positive (ZIKV titer ≥10), while 28 returned negative (ZIKV titer < 10), representing a false-positive rate of 30.4%.ConclusionAs this epidemic persists, data from this unique cohort of pregnant women with both local and travel-associated ZIKV exposure contributes to the growing knowledge base regarding implications of ZIKV in pregnancy.Disclosures All authors: No reported disclosures.