Clinical Case Study of ABO Hemolytic Disease in Full-term

Newborns with Positive Free Test and Direct Antiglobulin Test

Yan Jin

Department of Neonatology, the People’s Hospital of Baise, Baise, Guangxi,China

Keywords: Newborn, ABO Hemolytic dDisease, Free Antibody Test, Free Antibody Test, Direct Antiglobulin Test.

Abstract: ABO hemolytic disease of newborn (ABO-HDN) occurs early and progresses rapidly, which can lead to fetal

or neonatal anemia and neonatal hyperbilirubinemia. Severe intrauterine hemolysis can lead to fetal edema,

which can lead to fetal death or neonatal death. Severe or very severe hyperbilirubinemia can be complicated

with bilirubin encephalopathy, Affect the quality of life of newborns.In this study, full-term children with

ABO-HDN were divided into two groups according to the results of three serological tests of hemolysis.

Group A: single antibody release test (ART) was positive; Group B: ART positive, combined with at least

one hemolysis test positive in direct antiglobulin test (DAT) and free antibody test (FAT). The differences of

minimum hemoglobin, age at onset (h) and incidence of anemia between group A and group B were compared.

1 INTRODUCTION

ABO-HDN postpartum diagnosis methods mainly

rely on three hemolysis tests, as follows: ①FAT is

one of the necessary detection methods for the

postpartum diagnosis of ABO-HDN. It can check

whether there is free IgG antibody in the serum of the

newborn and indirectly indicate the concentration of

antibody in the serum of the child. It has guiding

significance for the hemolysis persistence of the child

and the clinical diagnosis and treatment of ABO-

HDN and the progress of the disease.In this test, adult

red blood cells of the same blood group are dropped

into the serum of sick newborns, and then anti human

globulin serum is dropped. Red blood cell

agglutination indicates that the test is positive. ②

DAT is a basic method for the diagnosis of ABO-

HDN. Positive enhancement of DAT is correlated

with the demand for phototherapy, and no false

positive DAT cases have been found (Valsami,

Politou, Boutsikou, et al.2015). DAT is a good

indicator for predicting the development of ABO-

HDN with ABO incompatibility (Aydin, Deveci,

Orman, et al.2016, Ulrich, Ellsworth, Carey 2014). If

there is IgG antibody inconsistent with erythrocyte

membrane surface antigen in newborns, it can

combine with erythrocyte membrane surface antigen

https://orcid.org/0000-0002-3465-7057

to form immune complex. However, due to the small

molecular weight of incomplete antibody, it can not

connect the antigen antibody complex well, which

only leads to the sensitization of red blood cells. After

the anti globulin serum is added, it can bind with the

incomplete antibody adsorbed on the red blood cells,

so as to link the sensitized red blood cells and produce

visible agglutination. Direct antiglobulin test (DAT)

can detect the presence of IgG anti-A or anti-B and

C3 on erythrocyte membrane to help identify

hemolysis as immune dependent or immune

independent. In the test, EDTA is best used for blood

samples (Shaz, Hillyer, Gil 2019). Meanwhile, the

neonatal DAT results can well predict their

compliance with the therapeutic indication of

hyperbilirubinemia(Shi, Ma, Zhu, et al 2018). ③

ART uses the reversibility of antigen antibody

reaction to release IgG antibody on sensitized red

blood cells of children with neonatal hemolytic

disease into normal saline through physical or

chemical methods. The released IgG antibody still

has biological activity. Standard type A or type B red

blood cells are added to the release solution, and the

released IgG antibody will sensitize red blood cells,

Subsequently, anti human globulin serum was added,

and the red blood cell aggregation visible to the naked

eye was positive. Among the three detection methods

Jin, Y.

Clinical Case Study of ABO Hemolytic Disease in Full-term Newborns with Positive Free Test and Direct Antiglobulin Test.

DOI: 10.5220/0011163000003444

In Proceedings of the 2nd Conference on Artiﬁcial Intelligence and Healthcare (CAIH 2021), pages 47-51

ISBN: 978-989-758-594-4

of abo-hdn hemolysis, the positive rate of antibody

release test is the highest(Daniel 2019). The

combination of heat release test and acid release test

can improve the positive detection rate of neonatal

ABO hemolytic disease and reduce the risk of missed

detection in release test (Chen, Deng, Huang, et

al.2019). Compare the results of slide test, tube test

and microcolumn gel test. Results the success rate of

ABO blood group identification by microcolumn gel

method was higher than that of slide method + test

tube method (You 2019).

Microcolumn gel cassette detection (MGCP) is

highly sensitive to serological tests of hemolytic

disease of the newborn by MGCP. It can significantly

improve the detection rate of ABO-HDN positive,

which can effectively reduce the missed diagnosis

rate. Moreover, the MGCP test is convenient and

quick, and easy to operate. It is better than the three

test of ABO-HDN in vitro with the test tube anti

human globulin test (TAT). Some studies also found

that compared with venous blood test, umbilical cord

blood hemolysis serological test is helpful to early

diagnose neonatal ABO-HDN and control the disease

as soon as possible (Hu, Zhang 2019). This study

retrospectively analyzed the clinical data of children

with ABO-HDN who met the diagnosis and treatment

criteria, and discussed the clinical manifestations and

influence of full-term ABO-HDN with positive free

test and direct antiglobulin test.

2 SUBJECTS AND METHODS

2.1 Study Subjects

The subjects selected 127 term neonates who met the

diagnostic criteria for the ABO-HDN.

2.2 Inclusion Criteria

According to the diagnostic criteria in practical

neonatology (4th Edition) for ABO-HDN of the

newborn, the diagnosis of ABO-HDN was confirmed

based on maternal and child ABO blood type (mother

with blood type O, child with blood type A or B, and

both mother and child with Rh blood type positive),

jaundice, and positive serological tests (DAT positive

or ART positive); Received phototherapy,

intravenous human immunoglobulin therapy;

Condition improved discharge; No specific treatment

was given out of hospital.

2.3 Exclusion Criteria

(1) Combined RH hemolytic disease; (2) Preterm

infants; (3) Children discharged automatically,

unable to implement treatment normally; (4)

Readmitted children; (5) Combined G-6-PD

deficiency; (6) Combined severe neonatal asphyxia;

(7) Complicated neonatal sepsis; (8) Children with

clinical symptoms of fever; (9) Comorbid congenital

heart disease.

2.4 Research Methods

SPSS statistics 16.0 software was used for statistical

analysis. The counting data were described by the

number of cases and percentage (n, %). The

measurement data were described by mean±standard

deviation (

SX ±

), and the differences between groups

were compared by independent sample t-test, analysis

of variance and chi square test; According to the

results of correlation factor analysis, the factors with

P<0.05 were included in logistic regression analysis,

and P<0.05 was statistically significant.

3 COMPARISON OF EFFECTS

OF DIFFERENT HEMOLYSIS

TEST GROUPS ON MINIMUM

HEMOGLOBIN, AGE AT

ONSET (H) AND INCIDENCE

OF ANEMIA

In this group of 127 cases, except ART positive, other

results of hemolysis tests were DAT positive in 18

cases (14.2%) and FAT positive in 64 cases (50.4%)

(see Table 1).

According to the different results of three

serological tests of hemolysis, they were divided into

two groups. Group A: ART positive alone; Group B:

ART positive, combined with at least one hemolysis

test of FAT and DAT positive. Independent sample t-

test was used between group A and group B to

compare the differences of minimum hemoglobin and

age at onset (h), and chi square test was used to

compare the differences of anemia incidence. The

results are shown in Table 2.

The lowest hemoglobin in group B was

significantly lower than that in group A (P<0.05), the

age at onset (h) was earlier than that in group A, the

difference was statistically significant (P=0.052), and

the incidence of anemia was higher than that in group

A (P<0.05).

CAIH 2021 - Conference on Artiﬁcial Intelligence and Healthcare

Table 1: Results of the three hemolysis tests in group A and

group B.

class,(n) ART

ositive

FAT

ositive

DAT

ositive

Group A

(

n=59

)

59 0 0

Group B

(

n=68

)

68 64 18

Table 2: The results of different hemolysis tests were

compared between the lowest hemoglobin, age (h) and

anemia.

class (n) Minimum

Hb(g/ L)

sx ±

Age at

disease

onset(h)

sx ±

Anemia

incidence2

(%)

Group A

(n=59)

147.39±

18.04

27.44±14.70 40.7%

Group B

(n=68)

130.74±

20.66

21.75±

17.53

72.1%

t value

2value

4.80 1.97 12.73*

P value 0.000 0.052 0.000

Note: * Use the chi-square test;1 x refers to the mean, s

refers to the standard deviation; 2 The denominator of the

incidence of anaemia was 127 term ABO-HDN cases

enrolled.

4 THE INFLUENCING FACTORS

OF MODERATE AND SEVERE

ANEMIA WERE ANALYZED BY

BINARY LOGISTIC

REGRESSION

4.1 Univariate Analysis of Moderate

and Severe Anemia in ABO-HDN

A total of 127 full-term ABO-HDN cases included in

the study were subjected to univariate analysis based

on the occurrence or absence of moderate to severe

anemia, using different hemolysis serology results as

independent variables, and the relevant assignments

are presented in table 3.

Table 3: Assignment table of relevant factors.

roject assignment

The hemolysis test

results were grouped

Group A =1

Group B =2

Table 4: Univariate analysis of moderate and severe anemia in term infants with ABO-HDN.

project classify No moderate or

severe anemia

occurred (n=104)

Moderate and

severe anemia

occurred (n=23)

χ2value P value

Different hemolysis

test results were

grouped

Group B 49 19 9.54 0.002*

Note: * P <0.05 has statistical differences

4.2 Binary Logistic Regression

Analysis of Moderate and Severe

Anemia in ABO-HDN

Table 4 shows that group B with ART positive and at

least one positive hemolysis test result of DAT and

FAT is P= 0.002, which is statistically significant (P

< 0.05). It is included in the binary logistic regression

analysis model, P < 0.05 is statistically significant,

and the results are shown in Table 5.

The results in Table 5 showed that the incidence

of moderate to severe anemia in ABO-HDN patients

with ART positive and at least one positive for DAT

and FAT was 5.33 times higher than that of ABO-

HDN patients with ART positive alone.

Table 5: Binary logistic regression analysis of related factors of moderate and severe anemia in ABO-HDN.

analytical factor B S.E. Wals P OR OR of 95% C.I.

lower

limit

superior

limit

Hemolysis test results

group B

1.67 0.58 8.21 0.004 5.33 1.70 16.75

Clinical Case Study of ABO Hemolytic Disease in Full-term Newborns with Positive Free Test and Direct Antiglobulin Test

According to the results of logistic regression, the

ROC curve was drawn. With the occurrence of

moderate and severe anemia as the reference, the area

under the ROC curve (AUC) of group B was 0.677,

indicating that there was a certain reference value for

predicting the occurrence of moderate and severe

anemia according to the results of hemolysis test. The

sensitivity of clinical prediction was 82.6% and the

specificity was 52.9%. It shows that the model has a

high accuracy in predicting the occurrence of

moderate and severe anemia (see Figure 1).

Figure 1: Logistic ROC Curve of regression Model.

5 CONCLUSIONS

In this study, the results of three postpartum

hemolysis tests for term ABO-HDN were divided

into two groups, A and B, to explore whether term

ABO-HDN with different positive serological test

results (ART, FAT, DAT) had differences. The

results showed that patients with ART positive,

combined with FAT and DAT at least one hemolysis

test positive group had significantly lower lowest

hemoglobin than the ART positive group alone, and

the results were statistically significant (P<0.05). It

was significantly earlier than the cases in the ART

positive group alone, and the result was statistically

significant (P < 0.05), and the incidence of anemia

was significantly higher than that in the ART positive

group alone (P < 0.05).

In the logistic regression analysis of the risk

factors for predicting the occurrence of moderate and

severe anemia, the case group with positive ART and

at least one of DAT and FAT was positive, compared

with the ART positive group alone, the risk of

moderate and severe anemia increased by 5.33 times.

The results were statistically significant

(P<0.05).DAT is a confirmed test of neonatal ABO

hemolysis, which has a lower positive rate in ABO

than in RH hemolysis, Although FAT cannot be used

as a decisive criterion for the diagnosis of neonatal

ABO hemolysis, it can detect whether there are free

IgG antibodies in neonatal serum and indirectly

reflect the degree and persistence of ABO, which is

of guiding significance for the clinical diagnosis and

treatment of ABO-HDN and disease

progress.Clinically, children with positive ABO-

HDN combined with at least one positive DAT and

FAT should be actively treated, so as to minimize the

incidence of children complicated with anaemia,

severe hyperbilirubinemia, and bilirubin

encephalopathy.In conclusion, in full-term ABO-

HDN cases, the serological hemolysis test was

positive for ART, combined with DAT and at least

one positive case with an increased moderate and

severe risk of FAT anemia.

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Clinical Case Study of ABO Hemolytic Disease in Full-term Newborns with Positive Free Test and Direct Antiglobulin Test