Development of pre-implantation genetic testing protocol for monogenic disorders (PGT-M) of Hb H disease.

Hb H disease is the most severe form of α-thalassemia compatible with post-natal life. Compound heterozygous α0-thalassemia- SEA deletion/α+-thalassemia- 3.7kb deletion is the commonest cause of Hb H disease in Thailand. Preimplantation genetics testing for monogenic disorders (PGT-M) is an alternative for couples at risk of the disorder to begin a pregnancy with a healthy baby. This study aims to develop a novel PCR protocol for PGT-M of Hb H disease- SEA/-3.7kb using multiplex fluorescent PCR. A novel set of primers for α+-thalassemia- 3.7kb deletion was developed and tested. The PCR protocol for α0-thalassemia- SEA deletion was combined for Hb H disease- SEA/-3.7kb genotyping. The PCR protocols were applied to genomic DNA extracted from subjects with different thalassemia genotypes and on whole genome amplification (WGA) products from clinical PGT-M cycles of the families at risk of Hb Bart's. The results were compared and discussed. The results showed three PCR products from α+-thalassemia- 3.7kb primer set, and three from α0thalassemiaSEA primer set. The results were consistent with the known thalassemia genotypes. The novel -α3.7 primers protocol was also tested on 37 WGA products from clinical PGT-M cycles giving accurate genotyping results and a satisfying amplification efficiency with the ADO rates of 2.7%, 0%, and 0% for HBA2, HBA1, and internal control fragments, respectively. This novel PCR protocol can precisely distinguish Hb H disease- SEA/-3.7kb from other genotypes. Additionally, this is the first PCR protocol for Hb H disease- SEA/-3.7kb which is optimal for PGT-M.

Change of contraceptive preference after the free-LARC program for Thai teenagers.

BACKGROUND: In 2014, the Thai government launched a free-of-charge long-acting reversible contraception (LARC) program for Thai female adolescents. However, its acceptance had not been reported. Therefore, this study aimed to describe contraceptive use among women of reproductive age before and after the program was implemented. METHODS: This retrospective cross-sectional study was carried out from the medical records of 9000 women of reproductive age, who attended the Family Planning Clinic at Maharaj Nakorn Chiang Mai Hospital between 2009 and 2018. The Chi-square test was used to compare the contraceptive methods administered before and after the program was implemented, and binary logistic regression was used to find the factors associated with implant use after completion of the program. RESULTS: Depot medroxy progesterone acetate (DMPA) injection was the most popular contraceptive method used among 40.9% of the women. The rates of subdermal implant use were increased significantly after the program was implemented (2.3-9.3%, p < 0.001). Implant use for adolescents aged less than 20 years increased from 2.6% to 56.4%, while DMPA was the most popular method used among adult women at 36.4%. Factors associated significantly with implant use after implement of the program included age of less than 20 years (aOR = 4.17 (CI: 1.84-9.44); p = 0.001) and nulliparity (aOR = 8.55 (CI: 3.77-19.39); p < 0.001). CONCLUSION: This study showed a significant increase in contraceptive implant use after the free-LARC program for adolescents had been applied. STATEMENT OF IMPLICATIONS: Contraceptive implant is the most effective hormonal reversible contraception. Its use rate is low among all age groups, but increased clearly after the free-of-charge program was applied for adolescents.

Pre-implantation genetic testing for Marfan syndrome using mini-sequencing.

Marfan syndrome (MFS1) is an autosomal dominant condition causing aortopathy including fatal aortic dissection. This study aimed to perform clinical PGT-M in a family with a history of MFS1 for two generations. A family with two members affected by MFS1 approached the hospital for PGT-M. The couple decided to join the project following extensive counselling and informed consent was obtained. The mutation contributory to MFS1 was identified using whole-exome sequencing (WES). A novel PGT-M protocol using multiplex fluorescent PCR and mini-sequencing was developed and tested. Ten blastocysts were subjected to PGT-M in two clinical PGT cycles. Mini-sequencing revealed four normal and six affected embryos. Microsatellite-based linkage analysis confirmed mutation analysis results in all samples. The embryos diagnosed as normal (non-MFS1) were chosen for transfer. A pregnancy was obtained in the third embryo transfer. Invasive prenatal diagnosis confirmed the normal genotype of the baby. This study demonstrated comprehensive management using the application of clinical-based diagnosis, WES for mutation identification within the MFS1 gene, mini-sequencing for embryo selection and microsatellite-based linkage analysis for backup of PGT-M results and contamination detection to assist couples in having a healthy child when there was a family history of Marfan syndrome.Impact StatementWhat is already known on this subject? Marfan syndrome (MFS1, OMIM#154700) is an autosomal dominant condition causing aortopathy including fatal aortic dissection. Pre-implantation genetic testing (PGT) is an alternative to traditional invasive prenatal diagnosis (PND) giving the couples the chance of starting pregnancy with the confidence that the baby will be unaffected. Most of the previous PGT reports employed microsatellite-based linkage analysis. A few PGT studies used sequencing, mini-sequencing and mutation analysis; however, the details of the techniques were not described.What do the results of this study add? Single-cell PCR protocol using multiplex fluorescent PCR and mini-sequencing was developed and validated. Two clinical PGTs cycles for Marfan syndrome were performed. A healthy baby was resulted. The details of multiplex fluorescent PCR and mini-sequencing protocols are described in this study so that the procedures can be reproduced.What are the implications of these findings for clinical practice and/or further research? Embryo selection can help the family suffering from Marfan syndrome for two generations to start a pregnancy with confidence that their child will be unaffected. This study also shows the use of a mini-sequencing protocol for PGT, which can be a universal protocol for other mutations by changing the PCR primers and mini-sequencing primers.

The successful strategy of comprehensive pre-implantation genetic testing for beta-thalassaemia-haemoglobin E disease and chromosome balance using karyomapping.

Thalassaemia is the commonest monogenic disease and causes a health and economic burden worldwide. Karyomapping can be used for pre-implantation genetic testing of monogenic disorders (PGT-M). This study applied karyomapping in two PGT-M cycles and made a comparison to polymerase chain reaction (PCR). Two families at risk of having beta-thalassaemia-haemoglobin E disease offspring decided to join the project and informed consent was obtained. Karyomapping results of family A (beta-thalassaemia (c.41_42delTCTT)-Hb E (c.26G>A) disease) revealed four normal, two beta-thalassaemia traits, one Hb E trait and six affected. Three embryos exhibited unbalanced chromosomes. One normal male embryo was transferred. Karyomapping results of family B (beta-thalassaemia (c.17A>T)-Hb E (c.26G>A) disease) revealed six Hb E traits and three affected. Three embryos were chromosomally unbalanced. One Hb E trait embryo was transferred. Two successful karyomapping PGT-M were performed, including deletion and single-base mutations. Karyomapping provides accuracy as regards the protocol and copy number variation which is common in pre-implantation embryos. Impact StatementWhat is already known on this subject? Thalassaemia syndrome is the commonest monogenic disease and causes a health and economic burden worldwide. Modern haplotyping using SNP array (aSNP) and karyomapping algorithms can be used for pre-implantation genetic testing of monogenic disorders (PGT-M). However, few clinical karyomapping PGT-M cycles have been done and validated so far.What do the results of this study add? Two successful clinical PGT-M cycles for beta-thalassaemia (c.41_42delTCTT and c.17A>T mutations)-haemoglobin E (c.26G>A) disease were performed using karyomapping. The outcome was two healthy babies. Multiplex fluorescent polymerase chain reaction (PCR) with mini-sequencing was also used for confirmation mutation analysis results. PCR confirmed haplotyping results in all embryos. Six embryos from both PGT-M cycles exhibited unbalanced chromosomes evidenced by aSNP.What are the implications of these findings for clinical practice and/or further research? Karyomapping provides accurate information quickly and the outcomes of the study will save time as regards protocol development, provide a usable universal PGT-M protocol and add additional copy number variation (CNV) information, chromosome number variation being a common issue in pre-implantation embryos.

A study of the association between angiotensinogen (AGT) gene polymorphism (M235T) and preeclampsia in Thai pregnant women.

AGT M235T gene polymorphism may cause increased blood pressure in preeclampsia in pregnancy; however, the evidence remains controversial. This study investigated the association between AGT M235T and preeclampsia in Thai pregnant women. A case-control study was conducted to compare the distributions of AGT M235T genotypes and alleles between 142 normotensive pregnancies as controls and 61 preeclampsia pregnancies as cases in a tertiary-care university hospital in Chiang Mai, Thailand. The results show that the distribution of AGT M235T genotypes (MM, MT and TT) of both groups were not significantly different (preeclampsia: 0.0, 16.4, 83.6%; control: 2.1, 22.5, 75.4%, respectively; p = .30). Additionally, there was no statistical difference in the distribution of AGT M235T alleles (M and T alleles) (preeclampsia: 8.2 and 91.8% versus control: 13.4 and 86.6%, respectively; p = .14). In this study, the distributions of AGT M235T were not different in both groups. Therefore, AGT M235T polymorphism may not play a significant role in preeclampsia pathophysiology in Thai population.Impact statementWhat is already known on this subject? Preeclampsia is one of the major complications during pregnancy; it significantly affects maternal and perinatal morbidity and mortality. Effort has been made to find markers and predictors that are associated with the pathophysiology of preeclampsia. AGT M235T gene polymorphism may cause increased blood pressure in preeclampsia pregnancy; however, evidences are still controversial.What do the results of this study add? We conducted a case-control study to compare the distributions of AGT M235T genotypes and alleles between 142 normotensive pregnancies as controls and 61 preeclampsia pregnancies as cases. The results show that preeclamptic women are more likely to deliver at an earlier gestational age and have a smaller baby in comparison with the normotensive group. In addition, women with preeclampsia had a higher chance of having an operative delivery and caesarean section. However, the distribution of AGT M235T polymorphism of preeclampsia women and the control group were not significantly different.What are the implications of these findings for clinical practice and/or further research? AGT M235T polymorphism may not play a significant role in preeclampsia pathophysiology in Thai population.

Comparisons of pregnancy outcomes between twin pregnancies with and without second-trimester amniocentesis.

OBJECTIVE: To assess the amniocentesis-related pregnancy loss rate and preterm birth rate among twin pregnancies undergoing amniocentesis. METHODS: A retrospective cohort study was conducted at a tertiary center. The study group included twin pregnancies undergoing amniocentesis during 16 to 22 weeks of gestation. The control group was those not undergoing amniocentesis. All amniocenteses were performed by the MFM specialists. The main outcomes were the rate of pregnancy loss (before 24 weeks) and preterm birth. RESULTS: A total of 332 cases in the study group and 1188 controls were analyzed. The percentages of maternal age ≥35 years, high parity, and cases complicated with medical diseases were significantly higher in the study group. The pregnancy loss rate after the procedure tended to be higher, but not significant, in the study group (3.0% vs 2.2% P = .383). Likewise, the rate of preterm birth in the study group was higher, but not significant (70.5% vs 66.0% P = .130). Logistic regression analysis to adjust confounding factors showed no significance of amniocentesis on pregnancy loss and preterm birth. CONCLUSION: Though amniocentesis in twin pregnancies has theoretical risk of pregnancy loss, it is relatively safe when performed by maternal-fetal medicine specialists. This information is useful for counseling, especially when performed by experienced hands.

The experience of genitourinary syndrome of menopause (GSM) among Thai postmenopausal women: the non-reporting issue.

Genitourinary syndrome of menopause (GSM) is common among postmenopausal women, but, in general, not all of the patients seek medical advice as this sensitive issue can cause them embarrassment. OBJECTIVES: To explore the prevalence of GSM among Thai postmenopausal women and their disclosure of and attitude towards GSM. METHODS: A questionnaire was used to obtain information on GSM from 499 Thai postmenopausal women who attended the Menopause Clinic at Maharaj Nakorn Chiang Mai Hospital, Chiang Mai, Thailand, from November 2015 to August 2016. RESULTS: The mean age of the 499 participants was 57.8 ± 7.2 years. It was notable that 87.2% of them had had GSM, and the prevalence increased with age. The most common symptoms were nocturia (77.7%) and vaginal dryness (51.7%). Among the symptomatic patients, 63.0% conveyed their problem to other people, i.e., friends and family, while 52.9% of them never reported to health care providers. The most common reason for not talking about their GSM was the acceptance of it being part of the natural aging process. CONCLUSIONS: GSM is common among Thai postmenopausal women. The prevalence of non-reporting is high and underreported.

Fetal Down syndrome screening models for developing countries; Part I: Performance of Maternal Serum Screening.

BACKGROUND: To identify the performance of fetal Down syndrome (DS) screening for developing countries. METHODS: A prospective study on MSS (maternal serum screening) with complete follow-ups (n = 41,924) was conducted in 32 network hospitals in the northern part of Thailand. Various models of MSS were tested for performance. RESULTS: MSS based on Caucasian reference range resulted in very high false positive rate (FPR; 13%) in our country, compared to the rate of 7.8% with our own (Thai) reference range, whereas the detection rate was comparable. As individual screening, C-S (contingent first trimester screening including PAPP-A, and free beta-hCG, classified as a) high risk [> 1:30], indicated for invasive diagnosis; b) intermediate risk [1:30-1500], indicated for STS; and c) low risk [< 1:1500], need no further tests.) was the most effective model (sensitivity 84.9%, FPR 7.7%) but nearly one-third needed the second trimester test (STS) because of intermediate results. Additionally, about one-third had their first visits in the second trimester and had no chance of FTS (first trimester screening). C-S plus STS had a sensitivity of 82.4% and FPR 8.1% whereas independent first and second trimester screening model (I-S) gave the sensitivity of 78.4% and FPR of 7.5% but was much more convenient and practical. CONCLUSION: C-S plus STS was the most effective models while I-S model was also effective and may be better for developing countries because of its simplicity and feasibility.

COVID-19 Vaccine Acceptance of Pregnant Women in Thailand.

Objective: This study investigated the COVID-19 vaccine acceptance rate among pregnant women in Thailand and explored factors influencing their willingness to receive the vaccine, to enhance vaccine uptake among hesitant pregnant women in the future. Methods: A prospective study was conducted at Maharaj Nakorn Chiang Mai Hospital, Chiang Mai, Thailand, in October 2022. The data was collected using face-to-face questionnaires comprising 29 closed-end questions. Pregnant women aged 18 years old or over visiting the antenatal care clinic were included. Results: The study included 200 participants, revealing a COVID-19 vaccine acceptance rate of 17%. Healthcare provider recommendations significantly increased vaccine acceptance by nearly two-fold (30.77%, p-value < 0.01). The major cause of vaccine hesitancy was the concern about vaccine safety that potentially harmed their babies (77.44%). Conclusion: The COVID-19 vaccine acceptance rate among pregnant women in Thailand was low. Healthcare provider recommendations played a pivotal role in positively impacting vaccine acceptance, highlighting their importance in increasing acceptance rates in the future.

This study investigated the COVID-19 vaccine acceptance rate among pregnant women in Thailand and explored factors influencing their willingness to receive the vaccine. The study included 200 participants, revealing a COVID-19 vaccine acceptance rate of 17%. Healthcare provider recommendations significantly increased vaccine acceptance by nearly two-fold (30.77%, p-value < 0.01). The major cause of vaccine hesitancy was the concern about vaccine safety that potentially harming their babies (77.44%). The COVID-19 vaccine acceptance rate among pregnant women in Thailand was low. Healthcare provider recommendations played a pivotal role in positively impacting vaccine acceptance, highlighting their importance in increasing acceptance rates in the future.

Effectiveness of the model for prenatal control of severe thalassemia.

OBJECTIVE: The aim of the research was to determine effectiveness of the model for prenatal control in reducing new cases of severe thalassemia. METHODS: Pregnant women at six tertiary centers were recruited to follow the model, consisting of (1) carrier screening using mean corpuscular volume (for alpha-thal-1 and beta-thal) and CMU-E screen (for HbE trait), (2) carrier diagnosis, (3) the couples at risk were counseled and offered prenatal diagnosis, and (4) termination of affected pregnancy. All neonates were evaluated for thalassemia. RESULTS: Of the 12,874 recruited pregnancies, 7008 were valid for analysis. Of them, 281 couples were identified to be at risk, Of the 281, 58 affected fetuses were identified and 55 pregnancies were terminated, whereas three did not accept pregnancy termination. All 6727 neonates at no risk were proven to be unaffected. The model had sensitivity and positive predictive value of 100% and 20%, respectively. The model could detect all of affected fetuses. CONCLUSION: The model could prenatally identify affected fetuses with a detection rate and negative predictive value of 100%. The model was highly effective to prenatally detect affected fetuses with an acceptable false positive rate.

Chorionic villous sampling: experience of 636 cases.

OBJECTIVE: To describe the technical aspects and complications of chorionic villous sampling (CVS). MATERIAL AND METHOD: The database of CVS procedures performed between January 2004 and August 2011 at Maharaj Nakorn Chiang Mai Hospital was assessed prospectively. Consecutive procedures during 10 to 14 gestational weeks were included into the present study. Indications, results, and complications of the CVS were extracted and analyzed. RESULTS: All 636 CVS procedures were successful. Indications for CVS were fetal karyotyping, DNA analysis for severe thalassemia disease, and for both in 36.5%, 50. 6%, and 12.9%, respectively However, 3.4% had inconclusive CVS result, and the second trimester prenatal diagnosis procedures needed to be done. There were five cases (0.8%) of fetal loss in the present study. Only two cases (0.3%) that fetal loss happened within two weeks of CVS procedures. Other minor complications such as vaginal bleeding and amniotic fluid leakage were found in 0.3 to 1.3% and had no long-term effect. No case with anomaly and procedure-related infection following the procedure was seen. CONCLUSION: CVS is a safe and reliable prenatal diagnosis procedure in the first trimester. In experienced operators, fetal loss rate was less than 1%. However a few cases will have inconclusive CVS results and may need a confirmation during the second trimester diagnostic procedures.

Effect of placenta penetration during cordocentesis at mid-pregnancy on fetal outcomes.

OBJECTIVES: To compare the rates of fetal loss, low birth weight, and preterm birth between pregnancies undergoing cordocentesis at mid-pregnancy with placenta penetration and those without it. METHODS: Consecutive cases of cordocenteses were prospectively recorded. The inclusion criteria for analysis were: (1) singleton pregnancies, (2) no fetal abnormalities, (3) gestational age of 18-22 weeks, and (4) procedures performed by experienced operators. The primary outcome was fetal loss rate, and the secondary outcomes were rates of failed procedures, low birth weight, and preterm birth. RESULTS: Of 6147 cordocenteses recorded, 2829 met the inclusion criteria with complete data for analysis. Of these, 654 procedures were further excluded because the puncture site was at cord insertion. The remaining 2175 cases, consisting of 615 procedures with placenta penetration and 1560 cases with no penetration, were analyzed. Cordocenteses with placenta penetration had a significantly higher rate of fetal loss (3.6% vs 1.3%, p = 0.01) as well as of low birth weight and preterm birth. CONCLUSION: Cordocentesis with placenta penetration carries a higher risk for fetal loss, preterm birth, and low birth weight. This information may be helpful in prenatal diagnosis counseling, and it may encourage performers to avoid placenta penetration, if possible.

Effect of umbilical cord bleeding following mid-pregnancy cordocentesis on pregnancy outcomes.

OBJECTIVES: To compare the adverse pregnancy outcomes among the three groups of umbilical cord bleeding following cordocentesis; a group of no bleeding (group 1), transient bleeding (group 2), and persistent bleeding (>60 s: group 3). METHODS: Consecutive cases of cordocentesis were prospectively recorded and followed up. The inclusion criteria for analysis consisted of (1) singleton pregnancies; (2) gestational age of 18-22 weeks; (3) no fetal anomalies; (4) no previous invasive procedures; (5) performed by experienced operators, and (6) known pregnancy outcomes. The main outcomes were rates of fetal loss, low birth weight and preterm birth. RESULTS: Of 2,174 procedures, 1,614 were in group 1, 509 were in group 2, and 51 were in group 3. The mean birth weight and gestational age at delivery were significantly lower in group 3 than those in groups 1 and 2. The fetal loss rate was significantly higher in group 3. Rates of low birth weight and preterm birth were significantly higher in group 3 whereas the rates in group 2 had a tendency to be higher than those in group 1. CONCLUSION: This study suggests that cord bleeding during cordocentesis carries a higher risk of fetal loss, low birth weight and preterm birth.

Preimplantation genetic diagnosis of alpha-thalassemia-SEA using novel multiplex fluorescent PCR.

PURPOSE: Preimplantation genetic diagnosis (PGD) is an alternative to prenatal diagnosis (PND) giving couples at risk a chance to start a pregnancy with a disease-free baby. This study aimed to develop a new PGD protocol for alpha-thalassemia(-SEA) mutation, the commonest Mendelian disorder. PATIENTS AND METHODS: Multiplex fluorescent PCR was employed for mutation, contamination and linkage analysis. A couple experienced termination of pregnancy following positive PND decided to join the project. RESULTS: Novel primers for alpha-thalassemia(-SEA) mutation amplifying 5 DNA fragments were developed. Two PGD cycles were performed, resulting in an un-affected baby. PND confirmed the heterozygous result. From 24 embryos, 87.5% of affected genotype were of best quality compared to 0% and 18.2% of those with normal and heterozygous, respectively. CONCLUSIONS: A novel PCR protocol for the common alpha-thalassemia(-SEA) mutation is reported. This test should be widely applicable. Interestingly, a potential effect of alpha-thalassemia(-SEA) mutation on preimplantation embryonic development was noticed.

Collagen Deposition and Inflammatory Response Associated with Macroporous Mesh Shrinkage in Incisional Hernia Repair: A Rat Model.

Background: Mesh repair is the current recommendation for the treatment of incisional hernia; however, the best mesh has yet to be determined. The objective of this study was to compare the inflammatory response and collagen deposition in primary incisional hernia repair (P) and different macroporous mesh materials, including polypropylene with poliglecaprone (PP-PG), polyvinylidene fluoride (PVDF), and polyester (PE), using quantitative methods. Methods: Sixty male rats were divided into four groups. Anterior abdominal wall defects were created and either suture or mesh repair was done. Rats were euthanized on days 14, 90, and 180, and the gross findings were recorded. The inflammatory and collagen levels in the abdominal wall tissues were measured using enzyme-linked immunosorbent assay (ELISA). Results: The PE group demonstrated significant mesh shrinkage at 180 days. The extent of PE mesh shrinkage ranged from 22-42% (mean = 30.49%). At 14 days, the PVDF group had higher interleukin-6 (IL-6) levels than the PP-PG (P = .004) and PE groups (P = .019). At 90 days, the collagen type I (Col I) levels in the PE group were significantly lower than those in the others, and the collagen type I/III (Col I/III) ratios in the PE group were lower than those in the P group (P = .006). Conclusions: The persistently high IL-6 levels until 180 days and the decrease in Col I levels and Col I/III ratio at 90 days seem to predict mesh shrinkage at 180 days. The mesh induces high Col I levels, but those associated with low Col III levels should be preferred.

Effect of cord puncture site in cordocentesis at mid-pregnancy on pregnancy outcomes.

OBJECTIVES: To compare the procedure and pregnancy outcomes of pregnancies undergoing cordocentesis between a group of cord insertion puncture and that of free loop puncture. MATERIALS AND METHODS: Pregnancies undergoing cordocenteses meeting the following inclusion criteria were recruited: (1) singleton pregnancies at gestational ages of 18-22 weeks; (2) no fetal abnormalities; (3) the procedures performed by experienced operators; and (4) the procedures of no placenta penetration. The recruited cases were classified into two groups: a group of cordocenteses with puncture site at placental cord insertion (Group 1) and those with puncture site at cord free loop (Group 2). The primary outcomes to be compared were rates of fetal loss, newborns with low birth weight and preterm birth. The secondary outcomes included a rate of successful procedures, umbilical cord bleeding, fetal bradycardia, maternal blood contamination, and duration of the procedure time. RESULTS: Of 6147 cordocenteses recruited, 2214 met inclusion criteria. The mean duration to complete the procedure in Group 1 was significantly shorter than that in Group 2 (4.5 ± 5.4 vs. 6.7 ± 8.5 min, Student's t-test; p = 0.001). The mean birth weight and gestational age at delivery were comparable between the two groups. No significant differences between the two groups were seen in rates of fetal loss, low birth weight, preterm birth, successful procedures, umbilical cord bleeding, or fetal bradycardia. However, the rate of maternal blood contamination was significantly higher in the group of puncture at the cord insertion site. CONCLUSION: Of cordocenteses performed based on accessibility and quality of visualization, cord insertion puncture may cause more maternal blood contamination but free loop puncture may be associated with more difficult procedures, while the safety was comparable.

Comparisons of outcomes after cordocentesis at mid-pregnancy between singleton and twin pregnancies.

OBJECTIVE: To compare the outcomes after mid-pregnancy cordocentesis between singleton and twin pregnancies. METHODS: We compared immediate complications and pregnancy outcomes between a cohort of twin pregnancies undergoing mid-pregnancy diagnostic cordocentesis between the years 1989 and 2010, and a control group of singleton pregnancies matched based on maternal age and years of the procedure with a ratio of 1:3. RESULTS: Among 6147 pregnancies undergoing cordocentesis during the study period, 122 twin fetuses met the study criteria and were matched with 336 singleton controls. Success rate of sampling was slightly higher in singleton than in twin gestations (98.8% vs 97.3%), and time required for the procedure was significantly longer in the twin group (8.1 vs 6.3 min, p = 0.02). Immediate complications, such as fetal bradycardia and transient bleeding from the puncture site, were significantly higher in twins (13% vs 6.0%, p = 0.001; and 34.8% vs 26.1%, p = 0.03, respectively). However, fetal loss rate within 2 weeks of cordocentesis was not different (1.4% and 1.1%, p = 0.42). CONCLUSION: Immediate complications following cordocentesis are significantly higher in twin pregnancies, but fetal loss rate within 2 weeks of the procedure is comparable.

Reference ranges of fetal aortic and pulmonary valve diameter derived by STIC from 14 to 40 weeks of gestation.

OBJECTIVE: To develop reference ranges of fetal aortic and pulmonary valve diameter derived from volume datasets of spatio-temporal image correlation (STIC). METHODS: A cross-sectional study was undertaken on low-risk pregnancies with well-established data from 14 to 40 weeks. Volume datasets of STIC were acquired for subsequent off-line analysis. Aortic and pulmonary valve diameters were measured in STIC multiplanar view using 4D-View version 9. Normal Z scores and centile reference ranges were constructed from these measurements against gestational age (GA) and biparietal diameter (BPD) as independent variables, using regression models for both mean and SD. RESULTS: A total of 606 volume datasets were successfully measured. Normal reference ranges for predicting mean values and SD of aortic and pulmonary valve diameter were constructed based on best-fit equations (linear function) as follows: mean aortic diameter (mm) was modeled as a function of GA (weeks) and BPD (mm) as - 2.4838 + 0.2702 × GA, (SD = 0.1482 + 0.0156 × GA) and - 1.5952 + 0.0989 × BPD (SD = 0.1672 + 0.00572 × BPD). Mean pulmonary diameter was modeled as - 2.5924 + 0.2935 × GA (SD = 0.2317 + 0.01524 × GA) and - 1.6830 + 0.1083 × BPD (SD = 0.1971 + 0.0059 × BPD). CONCLUSION: We have provided nomograms and Z scores of fetal aortic and pulmonary valve diameters. These reference ranges may be a useful tool in the assessment of fetal cardiac abnormalities.

Reference range of fetal splenic circumference from 14 to 40 weeks of gestation.

OBJECTIVE: To create a nomogram for fetal splenic circumference of normal fetuses. MATERIALS AND METHODS: A prospective, cross-sectional study was undertaken on normal pregnancies with certain date from 14 to 40 weeks of gestation. All fetuses were measured for fetal splenic circumference by tracing technique on transverse view of the fetal abdomen, using high-resolution real-time ultrasound with a 2-4 MHz convex transducer. RESULTS: A total of 684 normal pregnant women between 14 and 40 weeks of gestation were recruited. Fifty-eight were excluded because of poor image quality and fetal abnormality. The remaining 626 were available for analysis. Quadratic equation model was best fitted to estimate the 5th, 50th and 95th percentile range of splenic circumference at each gestational week. Fetal splenic circumference was gradually increased with gestational age with fitted equation as follows: splenic circumference (cm) = -4.181 + 0.456 (GA) -0.001 (GA)(2) (r = 0.942, p < 0.001). The table of nomogram for various percentile ranges was constructed. CONCLUSION: A normal reference range of fetal splenic circumference for each week of gestational age during 14-40 weeks is established.

Accuracy of immunochromatographic strip test in diagnosis of alpha-thalassemia-1 carrier.

OBJECTIVE: To determine the accuracy of alpha-thal immunochromatographic (IC) strip in diagnosis of alpha-thalassemia 1 carrier among pregnant women, using PCR for alpha-thalassemia 1 (SEA type) as a gold standard. MATERIAL AND METHOD: Asymptomatic pregnant women attending the antenatal care clinic were recruited Their blood samples were taken for IC Strip Test (alpha Thal IC strip, i+Med Laboratories Company Limited) in predicting alpha-thalassemia 1 carrier and separately sent for PCR for diagnosis of alpha-thalassemia 1 carrier as a gold standard RESULTS: Four hundred ninety nine pregnant women were recruited into the present study at various gestational weeks. The accuracy of alpha-Thal IC strip test was relatively high as shown in Table 1. Of them, 62 cases were proven to be alpha-thalassemia 1 trait and all ofthem had the results of positive IC strip, giving a sensitivity of 100%. However 45 pregnant women of non-alpha-thalassemia 1 trait had positive test, giving a specificity of 89%. CONCLUSION: The present study was solid evidence for clinical application of alpha-thal IC strip in screening program of thalassemia to reduce the need for PCR in diagnosis of a-thalassemia 1 carrier because of its very high sensitivity. The negative test reassures the non alpha-thalassemia 1 carrier status. Moreover, due to its simplicity, convenience to use, low cost, less-time consuming, clear interpretation and no need for either equipment or expensive laboratories, it may probably be very helpful in a massive screening program.