Objective To explore the clinical genetic counseling value of amniotic fluid cell culture and chromosome karyotype analysis in the second trimester of pregnancy. Methods The karyotype analysis results of 614 pregnant women who underwent amniocentesis in our hospital from December 2018 to March 2022 were analyzed retrospectively. Results 33 abnormal karyotypes were detected in 614 amniotic fluid samples, and the detection rate was 5.37%. Among the abnormal karyotypes, there were 21 cases with abnormal number (including chimera), 11 cases with abnormal structure, and 1 case with abnormal number and structure at the same time. The detection rates of abnormal karyotypes in seven types of prenatal diagnostic indications were 2.53%, 3.89%, 2.7%, 12.82%, 3.03%, 42.11% and 40% respectively. The detection rate of abnormal karyotype was the highest in the two indications of high risk of noninvasive DNA and chromosome abnormality of one spouse, and there was no significant difference between them; Indications with NT > 2.5mm ranked third; The detection rate of abnormal karyotype of the other four indications was the lowest, and there was no significant difference among the four indications. Conclusion Amniotic fluid cell culture and chromosomal karyotype analysis techniques are of great significance in preventing the birth of defective infants. Among the indications for Prenatal testing, the highest detection rate of amniotic fluid abnormal karyotypes is non-invasive high-risk and spouse chromosome abnormality indications